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• HOW TO CHOOSE THE RIGHT ELISA KIT

The enzyme-linked immunosorbent assay (ELISA or EIA) is a laboratory method to detect and quantify the presence of a protein in biological samples (1, 2).

When selecting an ELISA kit, researchers are often confronted with the question which assay to choose of the many commercially available kits.

It can be a challenge! Here are a few hints that may help.

HOW TO CHOOSE THE RIGHT ELISA KIT

As a general rule, before purchasing an assay, always read the protocol booklet (instructions for use – IFU or package insert) in detail. This should ensure that the kit will be suitable for your requirements. Check out the following:

1. ANALYTE

Which protein biomarker will you be measuring? Be sure to use the correct term during your search. Some biomarkers have alternative names (e.g. Sclerostin or SOST ELISA (SOST is actually the name for the gene that encodes Sclerostin).

2. SPECIES – SPECIFICITY – CROSS REACTIVITY

Verify if the assay can be used in the respective model such as e.g. human, rat, or mouse. Due to high homology between species, some ELISA kits work both in humans and in different animal species. As an example the biomarker ELISA kit for NT-proANP was developed for human use but due to the high sequence homology between species, the kit is successfully used to measure NT-proANP as a cardiac safety biomarker in various animal models (rat, mouse, rabbit, monkey).

3. SAMPLE TYPE

What is the sample type (matrix) you´ll be using (e.g. serum, EDTA-plasma, heparin-plasma, citrate-plasma, cell culture supernatants, urine..) ?

Verify if the assay is compatible for your sample type: check the package insert and, if available, check if there are validation data showing results (often found on the manufacturers website).

Of note: analysis of some biomarkers in the “wrong” matrix can lead to “false” results due to a matrix effect.

4. SAMPLE VOLUME

Check the amount of sample required per well (calculate volume to measure your samples in duplicates). Low sample volumes and precious samples are often a selection criterium.  

5. SENSITIVTY – BIOMARKER CONCENTRATIONS TO BE EXPECTED

Before choosing an assay, look into the validation data of the kit (often documented in the ELISA protocol booklet or in the validation data files).  

Reference values and pathological values in serum and/or plasma of the biomarker of interest are sometimes documented as well. These data can be helpful in selecting an appropriate assay. In some cases samples may require a pre-dilution. Therefore, always verify if the dilution buffer (assay buffer) is included in the kit or ask the assay developer for their input.

Of note: assays offering high sensitivity offer a different dynamic range than assays with a lower sensitivity. The dynamic range of an assay indicates the range of concentrations over which an assay can accurately quantify the analyte.

6. ASSAY PERFORMANCE – ASSAY VALIDATION

Careful evaluation of the assay´s performance characteristics is important in selecting an ELISA kit.

Choose an assay that has gone through a rigorous validation process. Check out if you can find data on the following performance characteristics:

• Accuracy- detection of a protein biomarker in clinical samples.
• Dilution linearity and parallelism  – recovery of the analyte of interest in diluted samples
• Specificity & cross-reactivity – making sure that you detect only the analyte of interest
• Precision – within-run and in-between run precision – ensuring precise and reproducible results within an across assay lots
• Calibration – ensures consistent performance over the range of the assay of the calibration curve
• Sample stability – ensures the stability of the analyte of interest (e.g. exposure of real samples to multiple freeze-thaw cycles, stability at room temperature..).

 

7. KIT COMPONENTS

Verify if the contents of the ELISA kit includes all the necessary components e.g. controls, assay dilution buffer. Consider storage requirements such as temperature sensitivity and expiration date.

8. CITATIONS & REFERENCES

Check if there are citations on the manufacturers website for the specific ELISA kit. Look into publications and seek feedback from researchers who have used the assay you are considering.

9. PRODUCT ORIGIN

Verify if the kit supplier is the kit manufacturer. More and more kits are repacked and are sold under different names, although it is always the same kit.

ELISA kit manufacturers will more likely give you qualified support as they “know” their product (e.g. availability of additional calibrators, controls, buffers.., technical know-how on the kit..).

10. CUSTOMER SUPPORT

Verify if the kit provider can provide timely and helpful customer service.

 

Related Literature

1.Enzyme-Linked Immunosorbent Assay: Types and Applications. Hayrapetyan H, Tran T, Tellez-Corrales E, Madiraju C. Methods Mol Biol. 2023;2612:1-17. doi: 10.1007/978-1-0716-2903-1_1. PMID: 36795355.

2. Enzyme Immunoassay (EIA)/Enzyme-Linked Immunosorbent Assay (ELISA). Lequin RM, Clinical Chemistry. 2005; 51, 12:  2415-2418.

 

February is marked as “Heart Month” drawing attention to heart disease and its prevention. Cardiovascular diseases (CVD) are the leading cause of death globally with an estimated toll of 17.9 million people per year (1).

Unhealthy diet, physical inactivity, tobacco use are only some of the  behavioral risk factors associated with heart disease. Health conditions that increase the risk include obesity, elevated blood sugar levels, high blood pressure, high low-density lipoprotein (LDL) cholesterol, and oxidative stress.

Biomarkers of Oxidative Stress in Heart Disease

What is oxidative stress?

 

Oxidative stress is caused by an imbalance between the production of free radicals (reactive oxygen species- ROS) and antioxidant defenses (endogenous antioxidant capacity). ROSs are generated as metabolic by-products in cells that can be stimulated by a variety of agents including pollutants, heavy metals, tobacco, drugs and many other. It has been postulated that oxidative stress can be responsible in the onset and progression of several diseases (e.g. cancer, diabetes, metabolic disorders, atherosclerosis, and cardiovascular diseases) (2).

Oxidized LDL (oxLDL) and Anti-oxidized LDL Autoantibodies

Accumulation of oxidized-low density lipoproteins (oxLDL) in the blood vessels triggers the onset of atherosclerosis which is a key element of the development of cardiovascular disease (CVD). The term atherosclerosis is of Greek origin, meaning thickening of the intimal layer of arteries and accumulation of fat (3). The process of atherosclerosis is initiated by LDL particles that lie in the sub-endothelial space of arteries (4). Thus, atherosclerosis is a disease of the vascular intima as a result of lipid oxidation. In the later stages of disease progression, plaques build up that rupture resulting in thrombosis (blockage of blood flow). oxLDL induces inflammatory responses in macrophages and dendritic cells (5).

Autoantibodies against oxidatively modified LDL (anti-oxLDL Ab)  have been detected in patients with atherosclerosis as well as in healthy individuals (6). It has been suggested that they mirror the occurrence of oxidation processes taking place in vivo. Anti-oxLDL Ab can be measured in human blood samples with a conventional ELISA assay (6-8). Numerous studies have implied that autoantibodies to oxLDL may allow improved CV risk stratification (6).

How can oxidative status / oxidative stress be measured?

 

Oxidative stress can be measured indirectly by different techniques. 

Example of a BIOMEDICA ELISA kit

 

1.MEASUREMENT OF ANTI-OXIDIZED LDL AUTOANTIBODIES 

ELISA for the detection of Anti-oxidized LDL Autoantibodies (oLAB ) | BI-20032

• Sample type: serum
• Sample volume 50µl/well
• Incubation time: 1.5 h / 30 min / 15 min
• Detection range: 0 – 1200 mU / ml
• Sensitivity: 48 mU / ml
• Precision: In-between-run (n=5): ≤ 8 % CV, Within-run (n=8): ≤ 4 % CV
• Use: Research use only
• Widely cited in over 70 publications

 

2.  MEASUREMENT OF BIOLOGICAL PEROXIDES 

Oxidative Stress Test – OXYSTAT Assay | BI-5007

Quick and easy assay to measure total peroxides in biological fluids.

• Method: colorimetric assay, 96 wells
• Sample type: serum, plasma, biological fluids
• Sample volume 10µl/well
• Assay time: 15 min
• Detection range: 0 – 660 µmol/l
• Sensitivity: 7 µmol/l
• Use: Research use only
• Widely cited in over 50 publications

 

Results show a direct correlation between free radicals and circulating biological peroxides and thus allow the characterization of the oxidative status in biological samples.

Literature

1. WHO-international health topics-cardiovascular diseases.
2. Reactive oxygen species in the vasculature: molecular and cellular mechanisms. Taniyama Y, Griendling KK. Hypertension. 2003 Dec;42(6):1075-81. doi: 10.1161/01.HYP.0000100443.09293.4F. Epub 2003 Oct 27. PMID: 14581295.
3. Atherosclerosis: process, indicators, risk factors and new hopes. Rafieian-Kopaei M, Setorki M, Doudi M, Baradaran A, Nasri H. Int J Prev Med. 2014 Aug;5(8):927-46. PMID: 25489440; PMCID: PMC4258672.
4. Oxidized LDL and anti-oxidized LDL antibodies in atherosclerosis – Novel insights and future directions in diagnosis and therapy. Hartley A, Haskard D, Khamis R. Trends Cardiovasc Med. 2019 Jan;29(1):22-26. doi: 10.1016/j.tcm.2018.05.010. Epub 2018 Jun 4. PMID: 29934015.
5. How Oxidized Low-Density Lipoprotein Activates Inflammatory Responses. Rhoads JP, Major AS. Crit Rev Immunol. 2018;38(4):333-342. doi: 10.1615/CritRevImmunol.2018026483. PMID: 30806246; PMCID: PMC6527110.
6. Anti-Oxidized LDL Antibodies and Coronary Artery Disease: A Systematic Review. van den Berg VJ, Vroegindewey MM, Kardys I, Boersma E, Haskard D, Hartley A, Khamis R. Antioxidants (Basel). 2019 Oct 15;8(10):484. doi: 10.3390/antiox8100484. PMID: 31618991; PMCID: PMC6826549
7. Circulating oxidized low density lipoprotein, autoantibodies against them and homocysteine serum levels in diagnosis and estimation of severity of coronary artery disease. Faviou E, Vourli G, Nounopoulos C, Zachari A, Dionyssiou-Asteriou A. Free Radic Res. 2005 Apr;39(4):419-29. doi: 10.1080/10715760500072156. PMID: 16028367.
8. Antibodies to oxidized low-density lipoprotein in patients following coronary artery revascularization. Miller ER 3rd, Erlinger TP, Blumenthal RS, Margolis S, Allen JK. Coron Artery Dis. 2003 Apr;14(2):163-9. doi: 10.1097/00019501-200304000-00009. PMID: 12655280.

 

 

 

 

Biomedica´s Osteoprotegerin (OPG) and Fibroblast Growth Factor (FGF23) assays were highlighted in the German Chronic Kidney Disease (CKD) cohort study to investigate the prognostic value of a total of nine mineral and bone disease biomarkers (MBD) (1). Brief, 4246 participants were enrolled and baseline serum samples of the MBD markers were prospectively observed for a median time of 6.5 years. The markers were compared to study the association of each biomarker with adverse cardiovascular (CV) outcomes and mortality. Of the nine biomarkers examined, Osteoprotegerin (OPG) best identified CKD patients who had the highest risk. The findings mirror the link between bone metabolism and CV disease in CKD patients and may help clinicians to identify patients who are at high risk for CV disease (1).

OPG a marker of cardiovascular mortality in CKD

The Kidney, Bone, and Heart Connection

 Chronic Kidney Disease (CKD), is a condition involving a gradual loss of kidney function. The kidneys lose their ability to filter toxins and extra fluid from the blood. There is no cure for this progressive disease and it is associated with a high morbidity and mortality rate that occurs especially in people with diabetes and hypertension (2). Worldwide, it is estimated that CKD is present in one out of ten adults (3).

Bone Disorders in Chronic Kidney Disease

 Other important roles of the kidney include maintaining bone health and balancing important minerals in the body. A majority of patients  with CKD have an elevated risk of developing disturbances of bone and mineral metabolism that lead to bone lesions (4). Thus, mineral an bone disorders (MBD) with biochemical and hormonal alterations are part of the complications associated with the progression of CKD (5).

 

Cardiovascular Events in Chronic Kidney Disease (CKD)

 Mineral and bone disorder in CKD can not only affect the bones but also other organs including the heart and blood vessels. It is well recognized that both atherosclerosis and arteriosclerosis are accelerated in patients with kidney failure (6). The high cardiovascular morbidity and mortality in CKD patients is mainly caused by progressive vascular calcification.

Osteoprotegerin (OPG) a Cardiovascular Risk Factor in CKD

 Osteoprotegerin (OPG) is a protein that regulates bone mass by inhibiting bone resorption. Apart from its traditional role in bone, OPG plays an important role in vascular disease and calcification as it interacts with endothelial and smooth muscle cells (7).

In a large prospective, population based study, severity, initiation, and progression of atherosclerosis was assessed in carotid arteries (8). The researchers demonstrated that OPG was an independent risk factor for the progression of atherosclerosis and onset of cardiovascular disease (CV) (8).

Serum OPG has also been shown to be a predictor of progression of atherosclerosis and coronary calcification in hemodialysis patients (9) and OPG serum levels increase with CKD disease progression and are associated with mortality in these patients (10). Moreover, high OPG levels are also associated with CV events in the general population as demonstrated in a large community based cohort (10). These data strongly reinforce OPG as marker for CV disease risk factor burden and predictor of CVD and mortality in the community (11).

 

Finally, the data from the recent German Chronic Kidney Disease cohort study investigating the prognostic value of nine mineral and bone disease associated biomarkers (MBD) showed that only serum OPG levels consistently identified the CKD patients who had the highest risk for adverse CV outcomes and mortality (1).

Osteoprotegerin (OPG) and Fibroblast Growth Factor (FGF23) can reliably be measured with a conventional ELISA assay

 Biomedica OPG ELISA (cat. no. BI-20403)

• Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
• Sample: Serum, plasma (EDTA, citrate, heparin)
• Sample volume: 20μl / well
• Detection range: 1.25-20 pmol/L (= 25 – 400 pg/mL)
• Sensitivity: 0.07 pmol/L (= 1.4 pg/mL)
• Incubation time: 4 h + 1 h + 30 min (room temperature)
• Precision: In-between-run (n=12): ≤ 5 % CV: Within-run (n=5): ≤ 3 % CV
• Widely cited in over 250 publications!

 

Biomedica FGF23 (C-terminal) ELISA (cat. no. BI-20702)

• Method: Sandwich ELISA, HRP/TMB, 12×8-well detachable strips
• Sample: Serum, plasma (EDTA, citrate, heparin)
• Sample volume: 50μl / well
• Detection range: 0.2 – 20 pmol/mL (= 1.54 – 150.4 pg/mL)
• Sensitivity: 0.08 pmol/L (= 0.6 pg/mL)
• Incubation time: 20-24 h  + 1 h+ 30 min (room temperature)
• Precision: In-between (n=10): ≤ 10 % CV; Within-run (n=6): ≤ 12 % CV

 

Publications

1. Association of mineral and bone biomarkers with adverse cardiovascular outcomes and mortality in the German Chronic Kidney Disease (GCKD) cohort. Reimer KC, Nadal J, Meiselbach H, Schmid M, Schultheiss UT, Kotsis F, Stockmann H, Friedrich N, Nauck M, Krane V, Eckardt KU, Schneider MP, Kramann R, Floege J, Saritas T; GCKD study investigators. Bone Res. 2023 Oct 20;11(1):52. doi: 10.1038/s41413-023-00291-8. PMID: 37857629; PMCID: PMC10587182.
2. Chronic kidney disease. Kalantar-Zadeh K, Jafar TH, Nitsch D, Neuen BL, Perkovic V. Lancet. 2021 Aug 28;398(10302):786-802. doi: 10.1016/S0140-6736(21)00519-5. Epub 2021 Jun 24. PMID: 34175022.
3. Prevalence, outcomes, and cost of chronic kidney disease in a contemporary population of 2·4 million patients from 11 countries: The CaReMe CKD study. Sundström J, Bodegard J, Bollmann A, Vervloet MG, Mark PB, Karasik A, Taveira-Gomes T, Botana M, Birkeland KI, Thuresson M, Jäger L, Sood MM, VanPottelbergh G, Tangri N; CaReMe CKD Investigators. Lancet Reg Health Eur. 2022. 20:100438. doi: 10.1016/j.lanepe.2022.100438. PMID: 36090671; PMCID: PMC9459126.
4. Chronic Kidney Disease-Mineral and Bone Disorder (CKD-MBD): Current Perspectives. Waziri B, Duarte R, Naicker S. Int J Nephrol Renovasc Dis. 2019 Dec 24;12:263-276. doi: 10.2147/IJNRD.S191156. PMID: 31920363; PMCID: PMC6935280.
5. Mineral Bone Disorders in Kidney Disease Patients: The Ever-Current Topic. Hu L, Napoletano A, Provenzano M, Garofalo C, Bini C, Comai G, La Manna G. Int J Mol Sci. 2022 Oct 13;23(20):12223. doi: 10.3390/ijms232012223. PMID: 36293076; PMCID: PMC9603742
6. Role of Chronic Kidney Disease (CKD)-Mineral and Bone Disorder (MBD) in the Pathogenesis of Cardiovascular Disease in CKD. Yamada S, Nakano T J Atheroscler Thromb. 2023 Aug 1;30(8):835-850. doi: 10.5551/jat.RV22006. Epub 2023 May 30. PMID: 37258233; PMCID: PMC10406631.
7. Role of crosstalk between endothelial cells and smooth muscle cells in vascular calcification in chronic kidney disease. Zhang YX, Tang RN, Wang LT, Liu BC. Cell Prolif. 2021. 54(3):e12980. doi: 10.1111/cpr.12980. Epub 2021 Jan 27. PMID: 33502070; PMCID: PMC7941222.
8. Osteoprotegerin is a risk factor for progressive atherosclerosis and cardiovascular disease. Kiechl S, Schett G, Wenning G, Redlich K, Oberhollenzer M, Mayr A, Santer P, Smolen J, Poewe W, Willeit J. Circulation. .109(18):2175-80. doi: 10.1161/01.CIR.0000127957.43874.BB. Epub 2004 Apr 26. PMID: 15117849.
9. Serum osteoprotegerin is a predictor of progression of atherosclerosis and coronary calcification in hemodialysis patients. Kurnatowska I, Grzelak P, Kaczmarska M, Stefańczyk L, Nowicki M. Nephron Clin Pract. 2011;117(4):c297-304. doi: 10.1159/000321169. Epub 2010 Sep 22. PMID: 20861651.
10. Circulating Osteoprotegerin in Chronic Kidney Disease and All-Cause Mortality. Kamińska J, Stopiński M, Mucha K, Pac M, Gołębiowski M, Niewczas MA, Pączek L, Foroncewicz B. Int J Gen Med. 2021 Jun 9;14:2413-2420. doi: 10.2147/IJGM.S302251. PMID: 34135625; PMCID: PMC8200134.
11. Biomarkers of the osteoprotegerin pathway: clinical correlates, subclinical disease, incident cardiovascular disease, and mortality.  Lieb W, Gona P, Larson MG, Massaro JM, Lipinska I, Keaney JF Jr, Rong J, Corey D, Hoffmann U, Fox CS, Vasan RS, Benjamin EJ, O’Donnell CJ, Kathiresan S. Arterioscler Thromb Vasc Biol. 2010 Sep;30(9):1849-54. doi: 10.1161/ATVBAHA.109.199661. Epub 2010 May 6. PMID: 20448212; PMCID: PMC3039214.
12. Association of mineral and bone biomarkers with adverse cardiovascular outcomes and mortality in the German Chronic Kidney Disease (GCKD) cohort. Reimer KC, Nadal J, Meiselbach H, Schmid M, Schultheiss UT, Kotsis F, Stockmann H, Friedrich N, Nauck M, Krane V, Eckardt KU, Schneider MP, Kramann R, Floege J, Saritas T; GCKD study investigators. Bone Res. 2023 Oct 20;11(1):52. doi: 10.1038/s41413-023-00291-8. PMID: 37857629; PMCID: PMC10587182.

 

 

 

 

Duchenne muscular dystrophy (DMD) is a hereditary neuromuscular disease that leads to progressive muscle fiber degeneration and weakness. There is no cure for this disease and current therapy consists on treatment with glycocorticoids (GC). GC therapy is linked to risk of bone loss and increased fracture risk. Despite their adverse effects GC remain the standard care to slow down disease progression (2).

Steroid therapy and fracture prevention in Duchenne Muscular Dystrophy

This recent study explored factors that are associated with incident fracture risk in glucocorticoid (GC)-treated patients with Duchenne muscular dystrophy (DMD): Risk Factors Associated with Incident Vertebral Fractures in Steroid-treated Males with Duchenne Muscular Dystrophy. Brief, vertical fractures (VF) were prospectively evaluated in 38 males with Duchenne muscular dystrophy at study baseline and 12 months . The authors concluded the following: ” The observation that ≥ 1 prevalent VF and/or non-VF were the strongest predictors of incident VFs at 12 months supports the need for prevention of first fractures in this high-risk setting. Bone age delay, a marker of GC exposure, may assist in the prioritization of patients in efforts to prevent first fractures.”

Steroid therapy and fracture prevention in Duchenne Muscular Dystrophy – The Biomedica IL-6 and Sclerostin ELISA were highlighted in this study.

High Sensitivity IL-6 ELISA  (cat. no. BI-IL6) – measurable values in serum and plasma samples.

• Format: 12×8 wells
• Sensitivity: 0.28 pg/ml
• Dynamic Range: 0-200 pg/ml
• Assay Time: 4 hours 30 minutes
• Sample Type: Serum, Plasma, Cell Culture Supernatants, Urine
• Sample Volume: 100 µl
• Alternative Name: Interleukin 6
• Kit includes 7 pre-diluted calibrators and 2 controls

 

Sclerostin ELISA (cat. no. BI-20492) 

• Format: 12×8 wells
• Sensitivity: 3.2 pmol/l (= 72 pg/ml)
• Dynamic Range: 0 to 240 pmol/l (=0-5400 pg/ml)
• Assay Time: 18-25h (overnight incubation) / 1 h /30 min
• Sample Type: Serum, Plasma, Cell Culture Supernatants, Urine
• Sample Volume: 20 µl
• Alternative Name: SOST
• Kit includes 6 pre-diluted calibrators and 1 control

 

Are you planning a study? Contact us to learn about our promotions info@bmgrp.com

Literature

1.Risk Factors Associated with Incident Vertebral Fractures in Steroid-treated Males with Duchenne Muscular Dystrophy. Phung K, McAdam L, Ma J, McMillan HJ, Jackowski S, Scharke M, Matzinger MA, Shenouda N, Koujok K, Jaremko JL, Wilson N, Walker S, Hartigan C, Khan N, Page M, Robinson ME, Saleh DS, Smit K, Rauch F, Siminoski K, Ward LM.J Clin Endocrinol Metab. 2023 Aug 23:dgad435. doi: 10.1210/clinem/dgad435. Epub ahead of print. PMID: 37610420.

Abstract

Purpose: Prevention of fractures is an unmet need in glucocorticoid (GC)-treated Duchenne muscular dystrophy. This study explored factors associated with incident vertebral fractures (VFs) to inform future fracture prevention efforts. Methods: VFs were evaluated prospectively at study baseline and 12 months on lateral spine radiographs in participants aged 4 to 25 years with Duchenne muscular dystrophy. Clinical factors were analyzed for their association with the change in Spinal Deformity Index (sum of the Genant-defined VF grades from T4 to L4) between baseline and 12 months. Results: Thirty-eight males were evaluated (mean ± SD age at baseline 11.0 ± 3.6 years; mean ± SD GC duration at baseline 4.1 ± 3.1 years; 74% ambulatory). Nine of 38 participants (24%) had 17 incident VFs, of which 3/17 VFs (18%) were moderate/severe. Participants with 12-month incident VF had lower mean ± SD baseline lumbar spine areal bone mineral density Z-scores (-2.9 ± 1.0 vs -1.9 ± 1.1; P = .049) and lower total body less head areal bone mineral density Z-scores (-3.1 ± 1.2 vs -1.6 ± 1.7; P = .036). Multivariable linear regression showed that at least 1 VF at baseline (P < .001), a higher number of antecedent non-VF (P < .001), and greater bone age delay at baseline (P = .027) were significant predictors of an increase in the Spinal Deformity Index from baseline to 12 months. Conclusion: The observation that ≥ 1 prevalent VF and/or non-VF were the strongest predictors of incident VFs at 12 months supports the need for prevention of first fractures in this high-risk setting. Bone age delay, a marker of GC exposure, may assist in the prioritization of patients in efforts to prevent first fractures.

2. Emerging therapies for Duchenne muscular dystrophy. Markati T, Oskoui M, Farrar MA, Duong T, Goemans N, Servais L. Lancet Neurol. 2022 Sep;21(9):814-829. doi: 10.1016/S1474-4422(22)00125-9. Epub 2022 Jul 15. PMID: 35850122.

 

 

 

BIOMEDICA provides custom analytical testing service for universities, biotechnology and pharmaceutical industries. Our skilled laboratory team conducts sample measurements for the quantification of biomarkers in various sample matrices. We deliver quality and reliable results tailored to your precise specifications. Our primary objective is to meet your specific goals! BIOMEDICA has a long track record in successfully conducting numerous research projects which have also resulted in publications in top journals.

BIOMEDICA Analytical Testing Service

ELISA ASSAYs for Biomarkers  DEVELOPED AND MANUFACTURED BY BIOMEDICA

We measure biomarkers using our propriety ELISA kits for the quantification of  analytes in serum, plasma, urine and cell culture supernatant. Check out our product portfolio 

ELISA ASSAYS FOR BIOMARKERS FROM OTHER MANUFACTURERS

We measure biomarkers in any kind of ELISA assay of your choice for any kind of analyte

LUMINEX TECHNOLOGY MULTIPLEX ASSAYS

We measure biomarkers using the Luminex xMAP^® (multiple analyte profiling) technology-based immunoassays that enable the simultaneous detection and quantification of multiple analytes.

NEXT-GENERATION SEQUENCING & RT-QPCR MICRORNA SERVICES

We provide a broad range of high-quality RNA services including RNA extraction, NGS and RT-qPCR and the custom analysis of microRNA signatures.

• Click here to learn more about the workflow

 

Related literature

Enzyme-Linked Immunosorbent Assay (ELISA). Tabatabaei MS, Ahmed M. Methods Mol Biol. 2022;2508:115-134. doi: 10.1007/978-1-0716-23763_10. PMID: 35737237.

Abstract

Enzyme-linked immunosorbent assay (ELISA) is one of the most specific and straightforward assays for detecting biomolecules in research and clinics. With advances in analytical methods, ELISA assay has been constantly optimized to improve its sensitivity, and different types of ELISA are now available to detect various biomarkers. This chapter provides an overall summary of the basic principle of ELISA, discusses different components of ELISA assay, and clearly outline protocols for different types of ELISA assays, including direct, indirect, sandwich, competitive, and nanoparticle-based ELISA.

Circulating miRNAs in bone health and disease. Grillari J, Mäkitie RE, Kocijan R, Haschka J, Vázquez DC, Semmelrock E, Hackl M. Bone. 2021 Apr;145:115787. doi: 10.1016/j.bone.2020.115787. Epub 2020 Dec 8. PMID: 33301964.

Abstract

microRNAs have evolved as important regulators of multiple biological pathways essential for bone homeostasis, and microRNA research has furthered our understanding of the mechanisms underlying bone health and disease. This knowledge, together with the finding that active or passive release of microRNAs from cells into the extracellular space enables minimal-invasive detection in biofluids (circulating miRNAs), motivated researchers to explore microRNAs as biomarkers in several pathologic conditions, including bone diseases. Thus, exploratory studies in cohorts representing different types of bone diseases have been performed. In this review, we first summarize important molecular basics of microRNA function and release and provide recommendations for best (pre-)analytical practices and documentation standards for circulating microRNA research required for generating high quality data and ensuring reproducibility of results. Secondly, we review how the genesis of bone-derived circulating microRNAs via release from osteoblasts and osteoclasts could contribute to the communication between these cells. Lastly, we summarize evidence from clinical research studies that have investigated the clinical utility of microRNAs as biomarkers in musculoskeletal disorders. While previous reviews have mainly focused on diagnosis of primary osteoporosis, we have also included studies exploring the utility of circulating microRNAs in monitoring anti-osteoporotic treatment and for diagnosis of other types of bone diseases, such as diabetic osteopathy, bone degradation in inflammatory diseases, and monogenetic bone diseases.

Preeclampsia is characterized by persistent high blood pressure and signs of liver or kidney damage that can affect pregnant women usually in the second half of pregnancy. It can also occur in women after the baby is delivered, mostly within 48 hours. Preelampsia occurs in around 2-8% of all pregnancies worldwide and is considered as a major cause of maternal and fetal mortality (1,2). It is responsible for over 70 000 maternal deaths and 500 000 fetal deaths worldwide every year (2). The cause of preeclampsia is still under debate and the clinical symptoms appear late during pregnancy (3).  Early diagnosis is essential to prevent morbidity and mortality that is associated with preeclampsia (4).

Endostatin a biomarker for early prediction of preeclampsia  

Endostatin is one of the most potent inhibitors of angiogenesis as it specifically inhibits the proliferation and migration of endothelial cells. Endostatin is a degradation product of collagen XVIII, a protein of the extracellular matrix (ECM) which is expressed in the basement membranes. Endostatin is a stable analyte that can reliably be measured in human blood and urine samples.

Biochemical markers may be useful to predict preeclampsia early, prior to the onset of clinical signs. A recent study explored the potential use of the protein biomarker Endostatin for the early prediction of preeclampsia (5).

BIOMEDICA human ENDOSTATIN ELISA │BI-20742 

• RELIABLE – validated according to FDA/ICH/EMEA guidelines
• FLEXIBLE  – only ELISA that can measure human Endostatin in serum, plasma and urine
• OPTIMIZED for clinical samples – wide dynamic range
• TRUSTED – cited and sample values provided  
• EFFICIENT – 20µl sample/well; results in 4.5 hours

 

BIOMEDICA mouse/rat ENDOSTATIN ELISA │BI-20742MR

• Fully validated for pre-clinical research (click here)
• Control included

 

Example of a BIOMEDICA ELISA assay kit

Literature

1. Preeclampsia diagnosis and management. Best Pract Res Clin Anaesthesiol. Overton E, Tobes D, Lee A. 2022 May;36(1):107-121. doi: 10.1016/j.bpa.2022.02.003. Epub 2022 Feb 10. PMID: 35659948.
2. Preeclampsia: Pathophysiology, Challenges, and Perspectives. Rana S, Lemoine E, Granger JP, Karumanchi SA. Circ Res. 2019 Mar 29;124(7):1094-1112. doi: 10.1161/CIRCRESAHA.118.313276. Erratum in: Circ Res. 2020 Jan 3;126(1):e8. PMID: 30920918.
3. The placenta and preeclampsia: villain or victim? Melchiorre K, Giorgione V, Thilaganathan B. Am J Obstet Gynecol. 2022 Feb;226(2S):S954-S962. doi: 10.1016/j.ajog.2020.10.024. Epub 2021 Mar 24. PMID: 33771361.
4. Early Detection of Preeclampsia Using Circulating Small non-coding RNA. Yoffe L, Gilam A, Yaron O, Polsky A, Farberov L, Syngelaki A, Nicolaides K, Hod M, Shomron N. Sci Rep. 2018 Feb 21;8(1):3401. doi: 10.1038/s41598-018-21604-6. PMID: 29467498; PMCID: PMC5821867.
5. Endostatin and Cystatin C as Potential Biomarkers for Early Prediction of Preeclampsia. Alshannag F, Zaki RMM, Hemida E, ElBakry MMM, Noureldeen AFH. ACS Omega. 2023 Nov 1;8(45):42776-42786. doi: 10.1021/acsomega.3c05586. PMID: 38024766; PMCID: PMC10652833.

 

Citations

•  Endostatin Is an Independent Risk Factor of Graft Loss after Kidney Transplant. Chu C, Hasan AA, Gaballa MMS, Zeng S, Xiong Y, Elitok S, Krämer BK, Hocher B. Am J Nephrol. 2020;51(5):373-380. doi: 10.1159/000507824. Epub 2020 Apr 22. PMID: 32320989.
• Validation of an enzyme-linked immunosorbent assay (ELISA) for quantification of endostatin levels in mice as a biomarker of developing glomerulonephritis. Wallwitz J, Aigner P, Gadermaier E, Bauer E, Casanova E, Bauer A, Stoiber D. PLoS One. 2019 Aug 12;14(8):e0220935. doi: 10.1371/journal.pone.0220935. PMID: 31404120; PMCID: PMC6690585.
• Higher Parathyroid Hormone Level Is Associated With Increased Arterial Stiffness in Type 1 Diabetes. Zobel EH, Theilade S, von Scholten BJ, Persson F, Tarnow L, Lajer M, Hansen TW, Rossing P. Diabetes Care. 2017 Mar;40(3):e32-e33. doi: 10.2337/dc16-2428. Epub 2017 Jan 6. PMID: 28062524.
• Endostatin in chronic kidney disease: Associations with inflammation, vascular abnormalities, cardiovascular events and survival. Kanbay M, Afsar B, Siriopol D, Unal HU, Karaman M, Saglam M, Gezer M, Taş A, Eyileten T, Guler AK, Aydin İ, Oguz Y, Tarim K, Covic A, Yilmaz MI. Eur J Intern Med. 2016 Sep;33:81-7. doi: 10.1016/j.ejim.2016.06.033. Epub 2016 Jul 7. PMID: 27394925.

 

 

In kidney transplantation recipients antibody mediated rejection (AMR) is a common complication that contributes to graft loss. C4d staining in kidney biopsies is widely used to identify the presence of complement activation in the small blood vessels of the kidney. Therefore C4d has been recognized as a marker for identifying AMR in kidney transplants (1).

C4d a biomarker of transplant rejection

 

C4d antibodies by Biomedica – for the identification of human complement split product C4d in paraffin and frozen sections as well as by flow cytometry.

Anti-C4d Antibody | BI-RC4D

• widely cited in over 90 citations
• for immunohistochemistry on paraffin embedded tissue and frozen sections
• use in kidney, heart, liver and other transplants

 

Anti-C4d Antibody (FITC) | BI-RC4D-FITC

• protocol for cell- or solid-phase bound C4 and C4d split product by flow cytometry
• for kidney, heart, liver and other transplants

 

The Biomedica C4d antibody was highlighted in the following publication: Response to treatment and long-term outcomes in kidney transplant recipients with acute T cell-mediated rejection. “Complement split product C4d staining was performed by immunochemical analysis of paraffin sections using polyclonal human anti‐C4d antibodies (Biomedica Gruppe, Vienna, Austria)”.

 

Literature

1. The Banff 2019 Kidney Meeting Report (I): Updates on and clarification of criteria for T cell- and antibody-mediated rejection. Loupy A, Haas M, Roufosse C, Naesens M, Adam B, Afrouzian M, Akalin E, Alachkar N, Bagnasco S, Becker JU, Cornell LD, Clahsen-van Groningen MC, Demetris AJ, Dragun D, Duong van Huyen JP, Farris AB, Fogo AB, Gibson IW, Glotz D, Gueguen J, Kikic Z, Kozakowski N, Kraus E, Lefaucheur C, Liapis H, Mannon RB, Montgomery RA, Nankivell BJ, Nickeleit V, Nickerson P, Rabant M, Racusen L, Randhawa P, Robin B, Rosales IA, Sapir-Pichhadze R, Schinstock CA, Seron D, Singh HK, Smith RN, Stegall MD, Zeevi A, Solez K, Colvin RB, Mengel M. Am J Transplant. 2020 Sep;20(9):2318-2331. doi: 10.1111/ajt.15898. Epub 2020 May 28. PMID: 32463180; PMCID: PMC7496245.

 

Bone Health & Osteoporosis – Biomarkers of Bone Regulation

Maintaining strong and healthy bones is essential for our well-being. Bone not only provides structural support for the body but also protects vital organs and serves as a provider for minerals such as calcium and phosphorus. Bone density and bone strength are key components of bone health.

Osteoporosis is a condition of weakened and fragile bones. It is the most common metabolic bone disease in the world (1) that can affect individuals of various ages, but is more commonly associated with aging. Both men and women can be affected, but postmenopausal women are at higher risk to develop Osteoporosis due to the decline of hormonal estrogen levels which plays a protective role in bone. Prevention of Osteoporosis includes a balanced diet and exercise.

Bone remodeling is a continuous process that is tightly regulated between bone resorption of old or damaged bone and the formation of new bone. Various hormones and factors are involved in bone metabolism. The bone cycle consists of different phases and markers of bone metabolism can be categorized as markers of bone formation, markers of bone resorption and markers of the regulation of  bone metabolism.

Bone Health & Osteoporosis – biomarkers of bone regulation

Protein biomarkers are often used in clinical research or clinical settings to assess bone health and to monitor the effectiveness of Osteoporosis treatments. Some of these biomarkers provide information about the regulatory processes involved in bone metabolism and turnover.

Monitoring these biomarkers can provide information on the overall health of bones. Some of these regulatory biomarkers include:

• Sclerostin (SOST) – is produced by osteocytes, bone cells embedded in the bone. Sclerostin is a bone specific Wnt pathway inhibitor, that negatively regulates bone formation, by promoting osteoclastogenesis and bone resorption (3). Elevated Sclerostin levels may indicate decreased bone formation.

 

• Dickkopf-1 (DKK-1) – is like Sclerostin an inhibitor of Wnt signaling, which is crucial for bone formation. Elevated serum DKK-1 promote bone resorption (4). DKK-1 levels may indicated suppressed bone formation.

 

• Receptor Activator of Nuclear Factor-Kappa B Ligand (RANKL) – is a key regulator of osteoclast activation and formation. RANKL promotes bone resorption by activating the bone osteoclasts. RANKL is secreted by osteocytes and is the most important factor of osteoclast formation (5).  

 

• Osteoprotegerin (OPG) – is the decoy receptor for RANKL. OPG regulates bone resorption (6). Changes in the OPG / RANKL ratio can affect bone remodeling.

 

• Fibroblast growth factor (FGF23) – is a hormone that regulates phosphate homeostasis and vitamin D metabolism. Abnormal FGF23 levels are associated with disorders affecting bone health e.g. hypophosphatemic rickets (7). 

   

  These protein biomarkers can easily be measured in human blood samples with an ELISA assay

   

  BIOMEDICA´s Bone Marker ELISA kits

  

• Sclerostin (SOST) ELISA  (BI-20492)

  • Most referenced Sclerostin ELISA +290 citations
  • Low sample volume – 20µl / well
  • Validation following international guidelines

   

• Bioactive Sclerostin ELISA (BI-20472)

  • Targets the receptor binding region
  • Rigorously validated for clinical samples according to FDA/ICH/EMEA guidelines
  • Low sample volume – 20 µl of serum/plasma per well 

   

• Dickkopf-1 (DKK-1) ELISA  (BI-20413)

  • Widely cited +170 publications
  • Direct measurement
  • Validated following international guidelines

   

• Osteoprotegerin (OPG) ELISA (BI-20403)

  • most referenced human OPG ELISA in +250 citations
  • day test, ready to use color coded reagents
  • controls included

   

• Free soluble RANKL ELISA (BI-20462)

  • Highly sensitive  – measurable concentrations in healthy subjects
  • Only ELISA that measures free, uncomplexed soluble RANKL
  • Cited in over +300 citations 

   

• FGF23 c-terminal multi-matrix ELISA (BI-20702)

  • for serum and plasma samples
  • full validation 
  • cited in +50 publications

   

• FGF23 intact ELISA (BI-20700)

  • for serum and plasma samples
  • full validation 
  • one-step ELISA

  

  Literature

  1. The clinician’s guide to prevention and treatment of osteoporosis. LeBoff MS et al., Osteoporos Int. 2022 Oct;33(10):2049-2102. doi: 10.1007/s00198-021-05900-y.
  2. Role of Wnt signaling and sclerostin in bone and as therapeutic targets in skeletal disorders. Marini F et al., Osteoporos Int. 2023 Feb;34(2):213-238. doi: 10.1007/s00198-022-06523-7.
  3. Inflammation and Bone Metabolism in Rheumatoid Arthritis: Molecular Mechanisms of Joint Destruction and Pharmacological Treatments. Maeda K et al., Int J Mol Sci. 2022 Mar 6;23(5):2871. doi: 10.3390/ijms23052871.
  4. Osteocyte-Related Cytokines Regulate Osteoclast Formation and Bone Resorption. Kitaura H et al., Int J Mol Sci. 2020 Jul 21;21(14):5169. doi: 10.3390/ijms21145169.
  5. The Role of Osteoprotegerin in Vascular Calcification and Bone Metabolism: The Basis for Developing New Therapeutics. Rochette L et al., Calcif Tissue Int. 2019 Sep;105(3):239-251. doi: 10.1007/s00223-019-00573-6.
  6. FGF23-related hypophosphatemic rickets/osteomalacia: diagnosis and new treatment. Fukumoto S. J Mol Endocrinol. 2021 Feb;66(2):R57-R65. doi: 10.1530/JME-20-0089.

   

 

It is well recognized that exercise has beneficial effects on overall health (1). Skeletal muscle has been described as a “secretory organ” that produces cytokines in response to muscle contraction (2). Thus, muscle communicates with various organs and tissues e.g. adipose tissue, liver, pancreas, bones, and brain (1).

IL-6 in exercise and health

Interleukin-6 (IL-6) is a cytokine that is primarily known to play a key role in inflammation and the regulation of the immune response. However, IL-6 is also an important signaling molecule during exercise. Numerous studies have demonstrated that Il-6 is produced locally in working skeletal muscle (3, 4).  Exercise, in particular intense or prolonged physical activity increases circulating IL-6 levels contributing to the regulation of glucose and lipid metabolism (4). Additional effects of IL-6 are discussed in Exercise and health — emerging roles of IL-6.

IL-6 ELISA, cat. no. BI-IL6

• HIGHLY SENSITIVE – measurable values in serum and plasma samples
• EASY – ready to use calibrators and controls
• RELIABLE – full validation package

Related ELISA kits

VEGF ELISA, cat. no. BI-VEGF

• SMALL SAMPLE VOLUME – 10µl/well
• HIGH SENSITIVITY – measurable values in both serum and plasma 
• RELIABLE –  rigorously validated according to FDA/ICH/EMEA guidelines
• EASY –  ready to use calibrators &  controls included

Angiopoietin-2 ELISA, cat. no. BI-ANG2

• Immediate results – assay range is optimized for clinical samples
• Characterized antibodies – detects all Angiopoietin-2 isoforms 
• Rigorously validated –  according to FDA/ICH/EMEA guidelines

Mouse/Rat Angiopoietin-2 ELISA, cat. no. BI-ANG2MR

• Low sample volume- 5µl
• Optimized for preclinical samples
• Sample values provided
• Detects all Angiopoetin-2 isoforms

Literature

1. Exercise and health — emerging roles of IL-6. Ellingsgaard H, Hojman P, Pedersen BK. Current Opinion in Physiology. 2019. 10; 49-54.
2. The effect of exercise on cytokines: implications for musculoskeletal health: a narrative review. Docherty, S., Harley, R., McAuley, J.J. et al. 2022. BMC Sports Sci Med Rehabil 14, 5.
3. IL-6 signaling in acute exercise and chronic training: Potential consequences for health and athletic performance. Nash D, Hughes MG, Butcher L, Aicheler R, Smith P, Cullen T, Webb R. Scand J Med Sci Sports. 2023. 33(1):4-19.
4. Muscle-derived interleukin-6: possible biological effects. Pedersen BK, Steensberg A, Schjerling P. J Physiol. 2001. 15;536(Pt 2):329-37.

Sepsis-induced brain dysfunction, referred to as sepsis-associated encephalopathy (SAE), is a common complication of sepsis affecting over half of sepsis patients in critical care units. It is linked to increased rates of mortality, morbidity, and long-term cognitive disability (1). The evaluation of SAE is crucial as it may be present in early stages of sepsis even before the diagnostic criteria for sepsis may be met (2). Biomarkers that are associated with brain injury could provide insights into the pathophysiological mechanisms of SAE and may be useful in patients for whom clinical assessment is challenging. C-type natriuretic peptide (CNP) and its amino-terminal propeptide NT-proCNP are natriuretic peptides, expressed in the vascular endothelium of many organs including the brain (3). Inflammatory mediators like IL-1 β and TNF-α can induce the release of NT-proCNP, suggesting a connection between systemic inflammation and the secretion and the regulation of NT-proCNP (4).

NT-proCNP in sepsis-induced brain dysfunction

The specific role of NT-proCNP as a biomarker for the assessment of SAE has been studied by Ehler et al., investigating NT-proCNP plasma concentrations in patients diagnosed with septic encephalopathy. The study revealed that highly elevated NT-proCNP levels in the early phase of sepsis might serve as an indicator for predicting the onset of SAE as the disease progresses (5).

NT-proCNP can reliably be measured in biological fluids with an ELISA assay from BIOMEDICA.

NT-proCNP ELISA (cat. no. BI-20823) kit highlights 

o   EFFICIENT – 20µl/well sample volume

o   CONVENIENT – 7 ready to use calibrators and 2 controls included

o   QUALITY – full validation following international quality guidelines

o   TRUSTED – widely cited

– Protocols available for urine, cell culture and non-human samples

Example of a Biomedica ELISA kit

 

MEASUREMENT OF NT-proCNP in NON-HUMAN SAMPLES

The sequence homology of NT-proCNP is very conserved among different species. We therefore assessed if the assay which is validated for human samples can be used in rat, mouse and pig samples. Other species types that share a high homology to human NT-proCNP can likely be measured with this assay (e.g. monkey, cats, dogs).  Learn more (NT-proCNP ELISA validation file pages 15-17). 

Literature

1. Septic-Associated Encephalopathy: a Comprehensive Review. Mazeraud A, Righy C, Bouchereau E, Benghanem S, Bozza FA, Sharshar T. Neurotherapeutics. 2020 Apr;17(2):392-403. doi: 10.1007/s13311-020-00862-1. PMID: 32378026; PMCID: PMC7283452.
2. Sepsis Associated Encephalopathy. Chaudhry N, Duggal AK.Adv Med. 2014;2014:762320. doi: 10.1155/2014/762320. Epub 2014 Sep 30. PMID: 26556425; PMCID: PMC4590973.
3. Prognostic value of circulating amino-terminal pro-C-type natriuretic peptide in critically ill patients. Koch A, Voigt S, Sanson E, Dückers H, Horn A, Zimmermann HW, Trautwein C, Tacke F.Crit Care. 2011;15(1):R45. doi: 10.1186/cc10007. Epub 2011 Jan 31. PMID: 21281508; PMCID: PMC3221974.
4. Inflammatory cytokines and NT-proCNP in Parkinson’s disease patients. Koziorowski D, Tomasiuk R, Szlufik S, Friedman A.Cytokine. 2012 Dec;60(3):762-6. doi: 10.1016/j.cyto.2012.07.030. Epub 2012 Aug 19. PMID: 22910321.
5. Diagnostic value of NT-proCNP compared to NSE and S100B in cerebrospinal fluid and plasma of patients with sepsis-associated encephalopathy. Ehler J, Saller T, Wittstock M, Rommer PS, Chappell D, Zwissler B, Grossmann A, Richter G, Reuter DA, Nöldge-Schomburg G, Sauer M. Neurosci Lett. 2019 Jan 23;692:167-173. doi: 10.1016/j.neulet.2018.11.014. Epub 2018 Nov 10. PMID: 30423400.

 

Cardiovascular toxicity (CV) plays a significant role in the failure of drug development. The use of blood-based biomarkers provides a way to detect CV toxicity during the preclinical stage, enabling the prioritization of compounds that show a lower risk inducing such adverse effects.

Cardiac Safety Biomarkers

The natriuretic peptides NT-proANP and NT-proBNP are cardiac hormones that are secreted when the heart muscle stretches. These peptides can be measured in blood samples and are well-known preclinical safety biomarkers used throughout drug development e.g NT-proBNP and NT-proANP – markers of drug-induced hypertrophy in rats (1-3).

BIOMEDICA provides robust ELISA assays for the accurate measurement of the these cardiac hormones for both human and rodent samples.

BIOMEDICA´s NT-proANP ELISA kit (BI-20892) has been independently validated in preclinical toxicology testing in rats:

• Cross-laboratory analytical validation of the cardiac biomarker NT-proANP in rat. 

 

In brief, the assay was evaluated by the Predictive Safety Testing Consortium, Cardiac Hypertrophy Working Group (PSTC—CHWG) in a cross-laboratory (5 laboratories) analytical assessment for use with rat samples. In summary, the assay has proven to be robust and technically adequate for the detection of NT-proANP serum levels in SD rats.

NT-proANP ELISA assay (cat. no. BI-20892)  

• 10 µl / well, serum or plasma
• widely cited as cardiovascular safety biomarker in rats
• for use in human and non-human samples (high cross-reactivity between species)

 

Rat NT-pro BNP ELISA assay (cat. no. BI-1204R) NEW

contact us for your special evaluation discount  info@bmgrp.com

• 10 µl / well,  serum or plasma
• kit control included
• sample values provided

 

Example of a Biomedica ELISA kit

 

PRODUCT INFORMATION

NT-proANP ELISA 

• Product code: BI-20892
• Method: ELISA
• Time to result: 3.5 hours
• Sample types: Serum, plasma, urine, cell culture supernatant (human, rat, mouse samples)
• Sample volume: 10 µl/well
• Sensitivity LOD: 0.05 nmol/l (= 0.64 ng/ml)
• Standard curve range: 0 – 10 nmol/l (= 0 – 127 ng/ml)
• Specificity: Endogenous and recombinant human NT-proANP (equivalent to proANP 1-98). Very high sequence homology between human and rodent (rat, mouse, and other species e.g. rabbit).
• NT-proANP ELISA assay is suitable for rat and mouse samples
• Instructions for use: click here

 

Rat NT-proBNP ELISA 

• Product code: BI-1204R
• Method: ELISA
• Time to result: 3.5 hours
• Sample types: rat serum and plasma
• Sample volume: 10 µl/well
• Sensitivity LOD: 21 pg/ml
• Standard curve range: 0 – 3200 pg/ml
• Specificity: Endogenous and recombinant rat NT-proBNP
• Instructions for use click here

 

Also available:

NT-proBNP human ELISA (cat. no. SK-1204), CE-marked, kit includes 2 controls 

CARDIAC SAFETY BIOMARKER ASSAYS in PRECLINICAL TOXICOLOGY TESTING 

References/Citations/Related Literature

All Citations on  NT-proANP as a cardiovascular safety biomarker in rat and mouse samples , click for all references

1. Cross-laboratory analytical validation of the cardiac biomarker NT-proANP in rat. Vinken P, Reagan WJ, Rodriguez LA, Buck WR, Lai-Zhang J, Goeminne N, Barbacci G, Liu R, King NM, Engle SK, Colton H.J Pharmacol Toxicol Methods. 2016; 77:58-65. doi: 10.1016/j.vascn.2015.10.002. PMID: 26516096.

2. Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium. Serum Natriuretic Peptides as Differential Biomarkers Allowing for the Distinction between Physiologic and Pathologic Left Ventricular Hypertrophy. Dunn ME et al., Toxicol Pathol. 2017; 45(2):344-352.
3. Natriuretic Peptides as Cardiovascular Safety Biomarkers in Rats: Comparison With Blood Pressure, Heart Rate, and Heart Weight. Engle SK, Watson DE.Toxicol Sci. 2016; 149(2):458-72. doi: 10.1093/toxsci/kfv240. Epub 2015 Nov 25. PMID: 26609138.

 

 

Type 2 diabetes mellitus (T2DM) is characterized by a persistent state of elevated blood sugar levels and glucose intolerance, resulting from the body´s incomplete response to insulin, accompanied by an increase in insulin production and a subsequent insulin deficiency. Individuals suffering from T2DM have an increased risk of cardiovascular disease (CVD). High glucose levels, insulin resistance, and chronic inflammation, contribute to endothelial dysfunction (ED) and atherosclerosis (1). ED refers to an impairment of the endothelium, the inner lining of blood vessels, which play an important role in regulating vascular health.

Sclerostin is associated with endothelial dysfunction in patients with type 2 diabetes

Sclerostin is a protein known primarily for its role in bone metabolism. It has also been identified of being linked to endothelial dysfunction in individuals diagnosed with type 2 diabetes (2). Sclerostin is predominantly secreted by osteocytes, cells that are embedded in the bone. However, vascular endothelial cells have also been observed to produce sclerostin leading to the discovery of its significant anti-calcifying role (3).

Sclerostin is associated with endothelial dysfunction in patients with type 2 diabetes: In an investigation in individuals with T2DM, researchers measured endothelial dysfunction by digital thermal monitoring (2). This method is a valid and noninvasive technique to evaluate endothelial function using temperature change on finger as a surrogate measure of the magnitude of vascular reactivity index (VRI) (4) . Serum Sclerostin levels were measured in the T2DM cohort with the Biomedica ELISA.  The prospective cross-sectional study revealed that serum sclerostin levels are positively associated with endothelial dysfunction measured in patients with T2DM.

A previous cross-sectional study in patients with T2DM, with/without cardiovascular disease, determined Sclerostin levels and its expression by RT-qPCR and immunohistochemistry in calcified and non-calcified artery of the lower limb from T2D. Serum Sclerostin was measured with an ELISA from Biomedica. Moreover, in vitro experiments were performed in vascular smooth muscle cells under calcifying conditions investigating the cardioprotective function of Sclerostin (5). The study provided evidence that supports the protective function of Sclerostin in the development of vascular calcification. The findings suggest that Sclerostin could potentially reduce the susceptibility to atherosclerosis by decreasing atherosclerotic plaque formation and underscore the significance of the bone-vascular axis when developing therapeutic strategies for treating impaired bone metabolism or vascular diseases (5).

Features and Benefits when measuring Sclerostin with the Biomedica ELISA kits

Sclerostin ELISA, #BI-20492

• TRUSTED – most referenced Sclerostin ELISA (+280 citations)
• LOW sample volume – 20µl sample /well
• For SERUM & PLASMA samples
• RELIABLE – fully validated  following international quality guidelines

 

 Bioactive Sclerostin ELISA, BI-20472

• CHARACTERIZED ANTIBODIES – targeting the receptor binding region
• EXTENSIVELY validated for clinical samples
• LOW sample volume – 20µl sample /well 

 

Literature

1. Endothelial Cell Dysfunction and the Pathobiology of Atherosclerosis. Garcia-de Los Ríos C, Medina-Casado M, Díaz-Chamorro A, Sierras-Jiménez M, Lardelli-Claret P, Sclerostin as a biomarker of cardiovascular risk in women with systemic lupus erythematosus. Cáliz-Cáliz R, Sabio JM. Sci Rep. 2022 Dec 14;12(1):21621. doi: 10.1038/s41598-022-25651-y. PMID: 36517533; PMCID: PMC9749620. Gimbrone MA Jr, García-Cardeña G. Circ Res. 2016 Feb 19;118(4):620-36. doi: 10.1161/CIRCRESAHA.115.306301. PMID: 26892962; PMCID: PMC4762052.
2. Serum sclerostin level is positively associated with endothelial dysfunction measured by digital thermal monitoring in patients with type 2 diabetes: A prospective cross-sectional study. Hsu BG, Wu DA, Yang HY, Chen MC.Medicine (Baltimore). 2023 Sep 8;102(36):e34649. doi: 10.1097/MD.0000000000034649. PMID: 37682176; PMCID: PMC10489308.
3. Sclerostin Protects Against Vascular Calcification Development in Mice. De Maré A, Opdebeeck B, Neven E, D’Haese PC, Verhulst A J Bone Miner Res. 2022 Apr;37(4):687-699. doi: 10.1002/jbmr.4503. Epub 2022 Feb 15. PMID: 35038187; PMCID: PMC9303214.
4. Digital thermal monitoring techniques to assess vascular reactivity following finger and brachial occlusions. Heath M, Gourley D, Naghavi M, Klies S, Tanaka H. J Clin Hypertens (Greenwich). 2020 Dec 7;23(1):122–7. doi: 10.1111/jch.14115. PMCID: PMC8030097.
5. Cardioprotective function of sclerostin by reducing calcium deposition, proliferation, and apoptosis in human vascular smooth muscle cells. González-Salvatierra S, García-Fontana C, Lacal J, Andújar-Vera F, Martínez-Heredia L, Sanabria-de la Torre R, Ferrer-Millán M, Moratalla-Aranda E, Muñoz-Torres M, García-Fontana B. Cardiovasc Diabetol. 2023 Nov 2;22(1):301. doi: 10.1186/s12933-023-02043-8. PMID: 37919715; PMCID: PMC10623848.

November is recognized as Diabetes Awareness Month, raising attention about diabetes and the importance of prevention and management of the disease that impacts millions of people worldwide.

Diabetes and Heart Disease – Risk stratification with NT-proBNP

Diabetes and heart disease are closely linked and patients suffering from diabetes have a 2- to 4 fold increased risk for developing cardiovascular diseases (CVD)  and concomitant heart failure (HF). Furthermore, HF patients with diabetes mellitus (DM) have a worse prognosis that those patients without DM (1, 2). In individuals with type 2 diabetes, high blood sugar levels and insulin resistance can lead to inflammation and atherosclerosis, a disease where arteries narrow, increasing the risk of CVD. Altered blood sugar levels are often associated with other CV risk factors, such as hypertension and obesity (3).

Diabetes and Heart Disease – Risk stratification with NT-proBNP 

Identifying patients who are at risk in developing HF remains a clinical challenge. Assessing the risk of HF development in patients with diabetes is important and the use of biomarkers could be beneficial (4). NT-proBNP, a protein that is specifically secreted by cardiac cells, is the “gold standard” biomarker for heart failure (5, 6). In patients with type 2 diabetes, NT-proBNP has been shown to predict death and cardiovascular events (7, 8). Moreover, a recent study investigated the relationships of NT-proBNP with the risk of developing diabetes and diabetes-related complication in initially healthy individuals (9). NT-pro BNP levels were measured in blood samples in a large cohort (European Prospecive Investigation Into Cancer and Nutrition) of over 27,000 individuals. The study demonstrated that NT-proBNP concentrations were inversely associated with the risk of type 2 diabetes in healthy individuals. Higher NT-proBNP levels are related to a higher risk of vascular disease in people who develop the disease. The authors conclude that NT-proBNP may be a promising biomarker to identify asymptomatic individuals who are at high risk developing diabetes independent of potential confounders.

NT-proBNP can be reliably measured in SERUM & PLASMA with a conventional ELISA assay.

NT-proBNP ELISA assay highlights (cat. no. SK-1204)

• RELIABLE – CE marked  –  for IVD use in the EU
• FLEXIBLE & EASY–  can be run in every lab
• CONVENIENT – Two controls included
• Assay independently validated in saliva samples
• Proficiency tested in accredited laboratory program Proficiency Testing Certificate 

 

Example of a BIOMEDICA ELISA assay test kit.

NT-proBNP ELISA (#SK-1204) click here for more information

Related products

NT-proANP ELISA (cat. no. BI-20892) – for human and rodent samples (10µl sample volume)

Rat NT-proBNP ELISA (cat. no. BI-1204R)  (10µl sample volume)

Literature

1. Epidemiology, Pathophysiology, Diagnosis and Treatment of Heart Failure in Diabetes. Park JJ. Diabetes Metab J. 2021 Mar;45(2):146-157. doi: 10.4093/dmj.2020.0282. Epub 2021 Mar 25. Erratum in: Diabetes Metab J. 2021 Sep;45(5):796. PMID: 33813813; PMCID: PMC8024162.
2. Diabetes as a risk factor for heart failure in women and men: a systematic review and meta-analysis of 47 cohorts including 12 million individuals. Ohkuma T, Komorita Y, Peters SAE, Woodward M. Diabetologia. 2019 Sep;62(9):1550-1560. doi: 10.1007/s00125-019-4926-x. Epub 2019 Jul 18. PMID: 31317230; PMCID: PMC6677875.
3. Diabetes leading to heart failure and heart failure leading to diabetes: epidemiological and clinical evidence. Palazzuoli A, Iacoviello M. Heart Fail Rev. 2023 May;28(3):585-596. doi: 10.1007/s10741-022-10238-6. Epub 2022 May 6. PMID: 35522391; PMCID: PMC10140137.
4. Assessing Cardiovascular Risk in Patients with Diabetes: An Update. Damaskos C, Garmpis N, Kollia P, Mitsiopoulos G, Barlampa D, Drosos A, Patsouras A, Gravvanis N, Antoniou V, Litos A, Diamantis E. Curr Cardiol Rev. 2020;16(4):266-274. doi: 10.2174/1573403X15666191111123622. PMID: 31713488; PMCID: PMC7903509.
5. NT-proBNP: The Gold Standard Biomarker in Heart Failure. McKie PM, Burnett JC Jr. J Am Coll Cardiol. 2016 Dec 6;68(22):2437-2439. doi: 10.1016/j.jacc.2016.10.001. PMID: 27908348.
6. 2022 AHA/ACC/HFSA Guideline for the Management of Heart Failure: Executive Summary: A Report of the American College of Cardiology/American Heart Association Joint Committee on Clinical Practice Guidelines. Heidenreich PA, Bozkurt B, Aguilar D, Allen LA, Byun JJ, Colvin MM, Deswal A, Drazner MH, Dunlay SM, Evers LR, Fang JC, Fedson SE, Fonarow GC, Hayek SS, Hernandez AF, Khazanie P, Kittleson MM, Lee CS, Link MS, Milano CA, Nnacheta LC, Sandhu AT, Stevenson LW, Vardeny O, Vest AR, Yancy CWCirculation. 2022 May 3;145(18):e876-e894. doi: 10.1161/CIR.0000000000001062. Epub 2022 Apr 1. PMID: 35363500.
7. NT-proBNP as a predictor of death and cardiovascular events in patients with type 2 diabetes. Malachias MVB, Wijkman MO, Bertoluci MC. Diabetol Metab Syndr. 2022 May 3;14(1):64. doi: 10.1186/s13098-022-00837-6. PMID: 35501909; PMCID: PMC9063067.
8. The role of NT-proBNP in the evaluation of diabetic patients with heart failure. Fringu FI, Sitar-Taut AV, Caloian B, Zdrenghea D, Comsa D, Gusetu G, Pop D. Acta Endocrinol (Buchar). 2020 Apr-Jun;16(2):183-191. doi: 10.4183/aeb.2020.183. PMID: 33029235; PMCID: PMC7535893.
9. Opposing Associations of NT-proBNP With Risks of Diabetes and Diabetes-Related Complications. Birukov A, Eichelmann F, Kuxhaus O, Polemiti E, Fritsche A, Wirth J, Boeing H, Weikert C, Schulze MB. Diabetes Care. 2020 Dec;43(12):2930-2937. doi: 10.2337/dc20-0553. Epub 2020 Aug 17. PMID: 32816995; PMCID: PMC7770272.

 

 

 

November is “Diabetes Awareness Month” raising attention to this fast growing and life-threating epidemic. Patients suffering from diabetes have a risk of additional health complications, including heart disease, strokes, and diabetic kidney disease (DKD). People who develop DKD mostly have few symptoms in the early stage of the disease, although the risk of developing severe kidney damage is very high. High blood sugar levels may damage the small blood vessels in the kidney leading to kidney damage, kidney failure, and high blood pressure (1). 

FGF23 and Sclerostin – novel biomarkers in diabetic kidney disease

Traditionally, the bone is regarded as a structural organ that gives the human body support and facilitates physical movement. However, bone is also a source of various hormones including fibroblast growth factor 23 and sclerostin that play an important role in regulating glucose metabolism and DKD (2).

FGF23 and Sclerostin  – novel biomarkers in diabetic kidney disease

 

FGF23 and Sclerostin – bone derived hormones regulate glucose metabolism

 

Fibroblast growth factor 23 (FGF23) is a bone-derived protein that regulates phosphate metabolism, by inhibiting renal phosphate reabsorption. There is increasing evidence that FGF23 plays a role in type 2 diabetes (T2DM), as FGF23 levels are elevated in these patients, even in individuals with preserved kidney function when compared to the general population (3). Phosphate independent effects on FGF23 following glucose loading were shown in a recent study demonstrating that FGF23 is associated with glucose, insulin and proinsulin levels, as well as obesity (4 ). Furthermore, FGF23 has also been shown to be associated with the development of gestational diabetes mellitus (5).

Sclerostin is a protein that is produced by bone cells that inhibits bone formation. Recent research suggests that Sclerostin also plays a role in lipid and glucose metabolism as serum sclerostin is negatively associated with insulin sensitivity as measured in obese, but not lean women (5). Sclerostin levels have also been shown to be increased in individuals with prediabetes (6).

FGF23 and Sclerostin can reliable by measured with conventional ELISA assays from BIOMEDICA.

FGF23 (C-terminal), #BI-20700 and FGF23 intact ELISA, #BI-20700

• RELIABLE – validated following international quality guidelines
• CITED in over 60 publications
• EASY – 8 standards and 2 controls included
• For SERUM & PLASMA samples
• HIGH QUALITY guaranteed!

 

Sclerostin ELISA, #BI-20492

• MOST referenced in more than 280 citations
• LOW sample volume – 20µl sample /well
• For SERUM & PLASMA samples
• RELIABLE – full validation package

 

Bioactive Sclerostin ELISA, BI-20472

• CHARACTERIZED ANTIBODIES – targeting the receptor binding region
• RIGOROUSLY validated for clinical samples
• LOW sample volume – 20µl sample /well

 

Literature

1. Renal microvascular disease predicts renal function in diabetes. Futrakul N, Futrakul P. Ren Fail. 2012;34(1):126-9. doi: 10.3109/0886022X.2011.623490. Epub 2011 Oct 20. PMID: 22010784.
2. The Emerging Role of Bone-Derived Hormones in Diabetes Mellitus and Diabetic Kidney Disease. Li Y, Gu Z, Wang J, Wang Y, Chen X, Dong B. Front Endocrinol (Lausanne). 2022 Jul 11;13:938830. doi: 10.3389/fendo.2022.938830. PMID: 35966090; PMCID: PMC9367194.
3. Earlier onset and greater severity of disordered mineral metabolism in diabetic patients with chronic kidney disease Wahl P., Xie H., Scialla J., Anderson C.A.M., Bellovich K., Brecklin C.et al.. (2012)  Diabetes Care 35, 994–1001 10.2337/dc11-2235 
4. Fibroblast Growth Factor 23, Glucose Homeostasis, and Incident Diabetes: Findings of 2 Cohort Studies, Amarens van der Vaart, Coby Eelderink, André P van Beek, Stephan J L Bakker, Peter R van Dijk, Martin H de Borst, The Journal of Clinical Endocrinology & Metabolism, Volume 108, Issue 10, October 2023, Pages e971–e978, https://doi.org/10.1210/clinem/dgad246.
5. Fibroblast growth factor 23 is associated with the development of gestational diabetes mellitus. Hocher CF, Chen X, Zuo J, Horvathova K, Hocher B, Krämer BK, Chu C. Diabetes Metab Res Rev. 2023 Aug 8:e3704. doi: 10.1002/dmrr.3704. Epub ahead of print. PMID: 37553983.
6. Serum sclerostin is negatively associated with insulin sensitivity in obese but not lean women. Aznou A, Meijer R, van Raalte D, den Heijer M, Heijboer A, de Jongh R. Endocr Connect. 2021 Feb;10(2):131-138. doi: 10.1530/EC-20-0535. PMID: 33480863; PMCID: PMC7983521.
7. Sclerostin and Insulin Resistance in Prediabetes: Evidence of a Cross Talk Between Bone and Glucose Metabolism. Daniele G, Winnier D, Mari A, Bruder J, Fourcaudot M, Pengou Z, Tripathy D, Jenkinson C, Folli F. Diabetes Care. 2015 Aug;38(8):1509-17. doi: 10.2337/dc14-2989. Epub 2015 Jun 17. PMID: 26084344.

Cigarette smoking is involved in the development of cardiovascular disease (CVD). It leads to changes in a number of biomarkers reflecting inflammation, oxidative stress, endothelial function, and lipid levels (1, 2) . A recent study investigated changes in biomarkers in individuals who stopped smoking (1). One of the biomarkers to measure oxidative stress in serum of individuals who quit smoking was “anti-oxidized low-density lipoprotein (anti-oxLDL) antibodies”. The anti-oxLDL antibodies were quantified with an assay from Biomedica.

Biomarkers of improved health after quitting smoking

Anti-Oxidized LDL Autoantibodies

Oxidized low-densitity lipoprotein (oxLDL) is believed to play a crucial role in in the development and progression of coronary artery disease (CAD) that involves atherosclerosis and inflammation. oxLDL in macrophages and smooth muscle cells accumulates and causes foam cell formation, which represents an early stage in the development of the disease. In fact, oxidized LDL appears to be around 24% higher in smokers compared to non-smokers (3). Autoantibodies targeting oxidatively modified LDL particles can serve as a parameter that consistently reflects the ongoing oxidation processes taking place in vivo. Studies demonstrate increased levels of autoantibodies against oxLDL in the bloodstream of individuals with coronary artery disease (4)

How can “Anti-Oxidized LDL Autoantibodies” be measured?

Autoantibodies targeting oxidatively modified LDL particles can easily be measured with a conventional ELISA assay.

Anti-Oxidized LDL Autoantibody ELISA (oLAB) assay (cat. no. BI-20032)

Highlights

• Widely cited
• Results in 2,5 hours
• 2 controls included
• Developed and manufactured by BIOMEDICA

 

Literature

1. Changes in biomarkers of endothelial function, oxidative stress, inflammation and lipids after smoking cessation: A cohort study. Colsoul ML et al., Eur J Clin Invest. 2023 Aug;53(8):e13996. doi: 10.1111/eci.13996. Epub 2023 Apr 8. PMID: 37000021.
2. Suitability of biomarkers of biological effects (BOBEs) for assessing the likelihood of reducing the tobacco related disease risk by new and innovative tobacco products: A literature review. Scherer G Regul Toxicol Pharmacol. 2018 Apr;94:203-233. doi: 10.1016/j.yrtph.2018.02.002. Epub 2018 Feb 9. PMID: 29428304.
3. Circulating oxidized low density lipoprotein, autoantibodies against them and homocysteine serum levels in diagnosis and estimation of severity of coronary artery disease. Faviou E et al., Free Radic Res. 2005 Apr;39(4):419-29. doi: 10.1080/10715760500072156. PMID: 16028367.
4. Biomarkers of improved health outcomes after smoking cessation. Neal L. Benowitz et al., Addiction Neuroscience,5: 2023, 100054,ISSN 2772-392. ISSN 2772-3925.

LRG – a reliable serum marker of disease activity in Inflammatory Bowel Disease: Inflammatory Bowel disease (IBD) like Crohn´s disease and ulcerative colitis affects millions of people worldwide. IBD is characterized by a chronic intestinal inflammation causing pain and swelling. The exact cause of IBD is not yet fully understood but it may involve the immune system, as well as genetic, and environmental factors. Research has shown that the immune system attacks harmless microorganisms or food in the gut that causes inflammation that leads to bowel injury.

Currently, the gold standard for assessing IBD disease activity are imaging tests, in particular endoscopy. The use of non-invasive biomarkers could be useful in clinical practice to monitor disease activity and to predict mucosal healing. CRP is often been used to evaluate IBD activity but research has shown that its specificity and low cut off value may not be ideal for use in clinical practice (1). Alternative biomarkers that more likely reflect intestinal inflammation are under investigation.

LRG – a reliable serum marker of disease activity in Inflammatory Bowel Disease

Leucine-rich α2 glycoprotein (LRG) is a protein that is produced by hepatocytes, neutrophils, and macrophages. It is a biomarker that is stimulated by inflammatory cytokines e.g. TNF-alpha, IL-1 beta, IL-11 and IL-6. However, studies have shown that release of LRG into the circulation does not depend on IL-6 and thus has a different mechanism from that of CRP. Therefore, LRG may better reflect intestinal inflammation better than LRG (3).

Recent studies have demonstrated that LRG is useful for predicting  mucosal healing and is a reliable serum biomarker for disease evaluation of clinical and endoscopic disease activity in individuals with inflammatory bowel disease (4-6).  

HOW CAN LRG BE MEASURED?

Leucine-rich α2 glycoprotein (LRG) can  easily and reliably be measured  in serum, plasma, urine and cell culture samples with a conventional ELISA (enzyme-linked immunosorbant assay) assay.

 

LRG ELISA – developed and manufactured by BIOMEDICA

• RELIABLE – validated following international quality guidelines (FDA, EMA, ICH) . The validation data can be found here.
• REFERENCE values provided for normal and pathological samples
• OPTIMIZED – assay range optimized for clinical samples, no additional testing required
• EASY – results in 3.5 h (protocol booklet)
• CONVENIENT – all reagents included; 7 pre-diluted standards/calibrators, 2 controls and sufficient assay buffer

 

Organizations for the study of IBD

• International Organization For the Study of Inflammatory Bowel Disease
• European Crohn’s and Colitis Organisation (ECCO)
• World Gastroenterology Organisation

 

Cited Literature

1. Leucine-rich alpha-2 glycoprotein as a marker of mucosal healing in inflammatory bowel disease. Yasutomi E, Inokuchi T, Hiraoka S, Takei K, Igawa S, Yamamoto S, Ohmori M, Oka S, Yamasaki Y, Kinugasa H, Takahara M, Harada K, Furukawa M, Itoshima K, Okada K, Otsuka F, Tanaka T, Mitsuhashi T, Kato J, Okada H. Sci Rep. 2021; 27;11(1):11086.
2.  Leucine-Rich Alpha-2 Glycoprotein Is a Reliable Serum Biomarker for Evaluating Clinical and Endoscopic Disease Activity in Inflammatory Bowel Disease. Shimoyama T, Yamamoto T, Yoshiyama S, Nishikawa R, Umegae S. Inflamm Bowel Dis. 2023; 29(9):1399-1408. 
3. Positioning and Usefulness of Biomarkers in Inflammatory Bowel Disease. Sakurai T, Saruta M. Digestion. 2023;104(1):30-41. 
4. Usefulness of Serum Leucine-rich Alpha 2 Glycoprotein in Crohn’s Disease: Is There Any Difference between Small Intestine and Colonic Lesions? Matsumoto S et al., Crohns Colitis 360. 2023; 23; 5(3):otad028.  Conclusions:  “LRG is more useful than CRP for predicting mucosal healing in patients with type L1. The superiority of LRG to CRP differs between small intestinal and colonic lesions.”
5. Leucine-Rich Alpha-2 Glycoprotein Is a Reliable Serum Biomarker for Evaluating Clinical and Endoscopic Disease Activity in Inflammatory Bowel Disease. Shimoyama T, Yamamoto T, Yoshiyama S, Nishikawa R, Umegae S.. Inflamm Bowel Dis. 2023; 29(9):1399-1408.  Conclusions: “Leucine-rich alpha-2 glycoprotein is a reliable serum biomarker for the assessment of clinical and endoscopic disease activity in patients with IBD. It can be an alternative to CRP for the assessment of UC.”
6. Leucine-rich alpha-2 glycoprotein is a potential biomarker to monitor disease activity in inflammatory bowel disease receiving adalimumab: PLANET study Shinzaki S et al., J Gastroenterol. 2021; 56(6):560-569. Conclusions: “Serum LRG is a useful biomarker of endoscopic activity both in CD and UC during the adalimumab treatment.”

 

Testing Cell Viability in Breast Cancer Cells with EZ4U – MTT Assay: In search for novel and effective drugs that don’t induce resistance or toxic effects the BIOMEDICA EZ4U – MTT Cell Viabilty Assay was used investigating various protein kinase C isoforms of Royleanones on the activity of breast cancer cell lines.

Royleanosones derive from plants from the genus Plectranthus with different varieties found throughout tropical Africa, Asia, and Australia. Various cytotoxic compounds with notable activities have been isolated from this plant. Learn more: Activity to Breast Cancer Cell Lines of Different Malignancy and Predicted Interaction with Protein Kinase C Isoforms of Royleanones.

Testing Cell Viability in Breast Cancer Cells with EZ4U – MTT Assay

In this study three natural Royleanones were tested for their inhibitory effects in three breast cancer cell lines namely: MCF-7 (high expression of estrogen receptors), SkBr3 (high expression of Her2^NEU ), and SUM159 (negative for both hormone receptors and for Her2).

In brief: the cell lines were grown at 37°C with 5% CO2. To test cell viability, cells were seeded at 10.000 cells per well and left for 24h. After cell attachment they were treated with various concentrations of the three natural Royleanones compounds and incubated for 24h. Cell viability was assayed with the EZ4U – MTT assay (Biomedica, Austria).

Short description: 20µl of the EZ4U substance (colorless non-toxic tetrazolium salts) was first added to each well. The mitochondria of the living cells oxidize the EZ4U substance to a yellow-colored formazan derivate. The color change is measured on a conventional plate reader at 450 nm using a reference wavelength of 620 nm.

EZ4U  Cell Viability & Cytotoxicity Assay (cat.no. BI-5000)

 

• Non-radioactive & non-toxic assay
• Reliable & Sensitive
• Convenient single-step incubation  – for use on living cells
• Widely cited in more than 250 publications

 

Find out more: 

• BROCHURE – EZ4U cell proliferation and cytotoxicity assay.
• Protocol Booklet

 

Citations using the BIOMEDICA EZ4U Assay (cat. no. BI-5000) in various cell-lines

Study on Anticancer Activity of 4, 4′-[1,4-phenylenebis(1,3,4-thiadiazole-5,2-diyl)] bis (azaneylylidene) bis (methaneylylidene) diphenolon Breast Cancer Cells. Saeed BMS et al., Arch Razi Inst. 2021; 76(4):821-827. 

Aberrant DNA Methylation, Expression, and Occurrence of Transcript Variants of the ABC Transporter ABCA7 in Breast Cancer. Zappe K et al., Cells. 2023;  24;12(11):1462. 

AQP3-Dependent PI3K/Akt Modulation in Breast Cancer Cells. Mlinarić M et al., Int J Mol Sci. 2023; 1;24(9):8133. 

Identification of F-Box/SPRY Domain-Containing Protein 1 (FBXO45) as a Prognostic Biomarker for TMPRSS2-ERG-Positive Primary Prostate Cancers. von Danwitz M et al.,  Cancers (Basel). 2023; 21;15(6):1890. 

Dual targeting of BCL-2 and MCL-1 in the presence of BAX breaks venetoclax resistance in human small cell lung cancer. Valko Z et al., Br J Cancer. 2023; 128(10):1850-1861. 

for more citations click here 

 

 

Periostin is a soluble extracellular matrix protein that plays a role in bone formation and has been linked to the development of various cancers, including breast cancer. It is secreted in the tumor environment mostly by cancer associated fibroblasts promoting the progression of cancer which includes cell survival, metastasis and also chemoresistance.

Periostin – a prognostic biomarker in breast cancer

The prognostic value of serum Periostin was investigated in a cohort of 509 non metastatic breast cancer patients. The results demonstrated the importance of Periostin in breast cancer and showed that serum Periostin is a prospective indicator for disease prognosis, regardless of the existence of micrometastases. Read more: High serum levels of periostin are associated with a poor survival in breast cancer.

Human serum Periostin levels were measured with the BIOMEDICA PERIOSTIN  ELISA.

Periostin ELISA (cat. no. BI-20433)

• • Low sample volume – 10 µl sample / well
  • For human serum/plasma/cell culture supernatants
  • High quality assay – validation following international guidelines
  • All reagents included – 7 ready to use calibrators and 2 controls
  • Numerous citations using the Biomedica Periostin ELISA

 

example of a BIOMEDICA ELISA assay kit with color-coded reagents, ready to use pre-diluted calibrators/standards and 2 (high/low) kit controls

 

The full characterization of the Periostin ELISA assay has been published in:

Characterization of a sandwich ELISA for the quantification of all human periostin isoforms. Gadermaier E et al., .J Clin Lab Anal. 2018, 32(2):e22252. doi: 10.1002/jcla.22252. 

Also available: Mouse Periostin ELISA (cat. no. BI-20433MS)

• Low sample volume – ≤ 5 µl / well
• For serum/plasma/cell culture supernatants

 

All ELISA kits are developed and manufactured by BIOMEDICA-Vienna/Austria

Related publications

High serum levels of periostin are associated with a poor survival in breast cancer. Rachner TD, Göbel A, Hoffmann O, Erdmann K, Kasimir-Bauer S, Breining D, Kimmig R, Hofbauer LC, Bittner AK. Breast Cancer Res Treat. 2020 Apr;180(2):515-524. doi: 10.1007/s10549-020-05570-0. Epub 2020 Feb 10. PMID: 32040688.

Periostin gene expression in neu-positive breast cancer cells is regulated by a FGFR signaling cross talk with TGFβ/PI3K/AKT pathways. Labrèche C, Cook DP, Abou-Hamad J, Pascoal J, Pryce BR, Al-Zahrani KN, Sabourin LA. Breast Cancer Res. 2021 Nov 22;23(1):107. doi: 10.1186/s13058-021-01487-8. PMID: 34809697; PMCID: PMC8607680.

Periostin Exon-21 Antibody Neutralization of Triple-Negative Breast Cancer Cell-Derived Periostin Regulates Tumor-Associated Macrophage Polarization and Angiogenesis. Fujikawa T, Sanada F, Taniyama Y, Shibata K, Katsuragi N, Koibuchi N, Akazawa K, Kanemoto Y, Kuroyanagi H, Shimazu K, Rakugi H, Morishita R.Cancers (Basel). 2021 Oct 11;13(20):5072. doi: 10.3390/cancers13205072. PMID: 34680221; PMCID: PMC8533925.

The Role of Extracellular Matrix Proteins in Breast Cancer. Lepucki A, Orlińska K, Mielczarek-Palacz A, Kabut J, Olczyk P, Komosińska-Vassev K. J Clin Med. 2022 Feb 25;11(5):1250. doi: 10.3390/jcm11051250. PMID: 35268340; PMCID: PMC8911242.

Fibroblast growth factor 23 (FGF23) is a protein that is produced by osteocytes, which are specialized cells found in bone tissue (1). FGF23 plays a key role in maintaining the balance of phosphate and vitamin D in the body (2). FGF23 primary function is to regulate blood phosphate levels by stimulating the removal of excess phosphate by the kidneys.

Elevated FGF23 levels in the bloodstream lead to a reduction in blood phosphate levels, a decrease in 1,25(OH)2 vitamin D, and the development of osteomalacia (3).

In addition to its known effects on bone and mineral regulation, it has been suggested that FGF23 may play a significant role in the development of type 2 diabetes mellitus (4). Findings showed that FGF23 is linked to indicators of obesity, metabolic syndrome and insulin levels.

FGF23 associated with the development of gestational diabetes

Gestational diabetes mellitus (GDM) is a type of diabetes that can develop during pregnancy in women who do not have existing diabetes. It is defined as glucose intolerance that is recognized first during pregnancy. GDM and impaired glucose tolerance during pregnancy are associated with persistent metabolic dysfunction three years after delivery, independent of other clinical risk factors (5).

FGF23 associated with the development of gestational diabetes

In the current study by Hocher CF and colleagues (6), the researchers investigated the association of maternal and foetal FGF23 with gestational diabetes mellitus (GDM) in a German birth cohort. Both FGF23 (c-terminal) and FGF23 intact levels were analysed in 826 random mother/child pairs with ELISA assays from BIOMEDICA. Of note: c-terminal FGF23 ELISA kits recognize both full-length intact FGF23 and c-terminal fragments of FGF23, and intact FGF23 assays solely detect full-length intact FGF23. The use of both assays in the current study is based on evidence from head to head comparison studies where clinical associations were reported based on the FGF23 assays used ( 7, 8). In summary, the study of a representative German birth cohort showed that maternal but not foetal iFGF23 is independently associated with GDM (6).

FGF23 can reliably be measured by ELISA assay

FGF23 intact ELISA, cat. no. BI-20700

FGF23 (c-terminal) ELISA,  cat. no.  BI-20702

BIOMEDICA´s FGF23 ELISA Kits:

• Low sample volume: 50µl / well
• For plasma (EDTA, citrate, heparın), serum, urine, cell culture supernatant
• Kit controls (high and low) included
• Full validation following international quality guidelines
• Numerous top journal citations

 

 

Literature:

1. The osteocyte: A multifunctional cell within the bone. Tresguerres FGF, Torres J, López-Quiles J, Hernández G, Vega JA, Tresguerres IF. Ann Anat. 2020 Jan;227:151422. doi: 10.1016/j.aanat.2019.151422. Epub 2019 Sep 26. Erratum in: Ann Anat. 2020 Jul;230:151510. PMID: 31563568.
2. Role of phosphate sensing in bone and mineral metabolism. Chande S, Bergwitz C. Nat Rev Endocrinol. 2018 Nov;14(11):637-655. doi: 10.1038/s41574-018-0076-3. PMID: 30218014; PMCID: PMC8607960.
3. FGF23 and Hypophosphatemic Rickets/Osteomalacia. Takashi Y, Kawanami D, Fukumoto S. Curr Osteoporos Rep. 2021 Dec;19(6):669-675. doi: 10.1007/s11914-021-00709-4. Epub 2021 Nov 10. PMID: 34755323.
4. The Emerging Role of Bone-Derived Hormones in Diabetes Mellitus and Diabetic Kidney Disease. Li Y, Gu Z, Wang J, Wang Y, Chen X, Dong B. Front Endocrinol (Lausanne). 2022 Jul 11;13:938830. doi: 10.3389/fendo.2022.938830. PMID: 35966090; PMCID: PMC9367194.
5. Gestational glucose tolerance and maternal metabolic profile at 3 years postpartum. Stuebe AM, Mantzoros C, Kleinman K, Gillman MW, Rifas-Shiman S, Seely EW, Rich-Edwards J. Obstet Gynecol. 2011 Nov;118(5):1065-1073. doi: 10.1097/AOG.0b013e3182325f5a. PMID: 22015874; PMCID: PMC3268071.
6. Fibroblast growth factor 23 is associated with the development of gestational diabetes mellitus. Hocher CF, Chen X, Zuo J, Horvathova K, Hocher B, Krämer BK, Chu C. Diabetes Metab Res Rev. 2023 Aug 8:e3704. doi: 10.1002/dmrr.3704. Epub ahead of print. PMID: 37553983.
7. C-terminal and intact FGF23 in kidney transplant recipients and their associations with overall graft survival. Chu C, Elitok S, Zeng S, Xiong Y, Hocher CF, Hasan AA, Krämer BK, Hocher B. BMC Nephrol. 2021 Apr 8;22(1):125. doi: 10.1186/s12882-021-02329-7. PMID: 33832449; PMCID: PMC8033679.
8. Fibroblast growth factor 23 concentrations and modifying factors in children from age 12 to 24 months. Enlund-Cerullo M, Hauta-Alus H, Valkama S, Rosendahl J, Andersson S, Mäkitie O, Holmlund-Suila E. Bone. 2020 Dec;141:115629. doi: 10.1016/j.bone.2020.115629. Epub 2020 Sep 10. PMID: 32919110.

 

 

Breast cancer is the most common detected cancer worldwide with 2.3 million new cases and 685, 000 deaths in 2020 (1).

1 in 8 women are predicted to be diagnosed with breast cancer in their lifetime. 

Early detection of breast cancer is the key to successful outcomes.

Breast Cancer Awareness Month – October 2023

Find out more about some of the different breast cancer associations worldwide and learn about what you should be aware of:

• National Breast Cancer Foundation Inc
• WHO Europe
• Breast Cancer UK
• Breast Cancer Foundation New Zealand
• Canadian Cancer Society
• Africa Health Organisation
• American Association for Cancer Research
• Japanese Breast Cancer Society

 

BIOMEDICA – Biomarker ELISA Kits for Breast Cancer Research

PERIOSTIN ▪ NEUROPILIN-1 ▪ OSTEPROTEGERIN ▪ soluble RANKL ▪ SEMAPHORIN 4D

check out our Brochure Biomarker ELISAs in Oncology

Assay Kit Highlights

√  EASY –ready to use calibrators & controls included (color-coded reagents)
√  FULL VALIDATION PACKAGE – assays are optimized for clinical samples
√   HIGH QUALITY GUARANTEED – results you can rely on
√  WIDELY CITED in 1500 + publications
√  COMPETENT CUSTOMER SERVICE

 

BIOMEDICA – Complete ready-to-use ELISA kits for superior performance and reproducibility

Literature

1. Current and future burden of breast cancer: Global statistics for 2020 and 2040. Arnold M, Morgan E, Rumgay H, Mafra A, Singh D, Laversanne M, Vignat J, Gralow JR, Cardoso F, Siesling S, Soerjomataram I Breast. 2022 Dec;66:15-23. doi: 10.1016/j.breast.2022.08.010. Epub 2022 Sep 2. PMID: 36084384; PMCID: PMC9465273.
2. The prevention and treatment of breast cancer- related lymphedema: A review. McEvoy MP, Gomberawalla A, Smith M, Boccardo FM, Holmes D, Djohan R, Thiruchelvam P, Klimberg S, Dietz J, Feldman S. Front Oncol. 2022 Dec 6;12:1062472. doi: 10.3389/fonc.2022.1062472. PMID: 36561522; PMCID: PMC9763870.
3. Breast Cancer in India: Screening, Detection, and Management. Bhardwaj PV, Dulala R, Rajappa S, Loke C. Hematol Oncol Clin North Am. 2023 Jun 15:S0889-8588(23)00069-2. doi: 10.1016/j.hoc.2023.05.014. Epub ahead of print. PMID: 37330342.
4. Outcomes in breast cancer-does ethnicity matter? Yap YS ESMO Open. 2023 Jun;8(3):101564. doi: 10.1016/j.esmoop.2023.101564. Epub 2023 Jun 7. PMID: 37290358; PMCID: PMC10265619.

 

 

Ovarian cancer ranks as the primary contributor of death in patients with gynecological malignancies and over 70% of patients receive their diagnosis at an advanced stage.  Despite advances in treatment, a large number of ovarian cancer patients still are confronted with a poor overall prognosis. The discovery of blood-based biomarkers that can predict outcomes or provide prognostic insights is promising as findings could lead to the identification of novel drug targets and tailored treatments for ovarian cancer patients.

Neuropilin-1 is a blood-based prognostic biomarker in patients with ovarian cancer

Neuropilin-1 is a transmembrane cell-surface receptor that has been implicated in various aspects of cancer biology. It is found on the surface of cancer cells and is shed into the circulation where it can successfully be measured in blood samples with a conventional ELISA assay. Neuropilin-1 is associated with tumor progression and the formation of new blood vessels to support tumor growth is a process called angiogenesis. Neuropilin-1 interacts with molecules like vascular endothelial growth factor (VEGF) and Semaphorins promoting the survival and migration of cancer cells.

Neuropilin-1 is a blood-based prognostic biomarker in patients with ovarian cancer

The BIOMEDICA Neuropilin-1 (NRP-1) ELISA assay was highlighted in a recent study (1) showing for the first time that NRP-1 is a blood based prognostic biomarker in patients with ovarian cancer. The results encourage further research to analyse whether soluble NRP-1 could be considered as a predictive and prognostic routine tool in patients with ovarian cancer.

EASY measurement of soluble Neuropilin-1 in blood samples:

Total Soluble Neuropilin-1 ELISA | BI-20409

• Method: conventional 96-well ELISA format
• Sample volume: 10µl / well
• Sample matrix: human serum, plasma, urine, cell culture supernatant
• High homology between human NRP-1 and non-human samples –  protocol available
• Full validation package – click here 

 

Only assay that measures free and ligand bound soluble NRP1

Reference

1. Clinical impact of soluble Neuropilin-1 in ovarian cancer patients and its association with its circulating ligands of the HGF/c-MET axis.   Klotz DM, Kuhlmann JD, Link T, Goeckenjan M, Hofbauer LC, Göbel A, Rachner TD, Wimberger P. Front Oncol. 2022 Oct 21;12:974885. doi: 10.3389/fonc.2022.974885. PMID: 36338759; PMCID: PMC9635484.

 

Abstract

Background: Neuropilin (NRP) is a transmembrane protein, which has been shown to be a pro-angiogenic mediator and implicated as a potential driver of cancer progression. NRP-1 up-regulation in ovarian cancer tissue predicts poor prognosis. However, the clinical relevance of the soluble form of NRP-1 (sNRP-1) as a circulating biomarker in ovarian cancer patients is unknown.

Methods/patients cohort: sNRP-1 levels were quantified in a cohort of 88 clinically documented ovarian cancer patients by a commercially available sNRP-1 enzyme-linked immunosorbent assay (ELISA) kit (Biomedica, Vienna, Austria). Patients (81.8% with FIGOIII/IV) received primary cytoreductive surgery with the aim of macroscopic complete resection (achieved in 55.7% of patients) and the recommendation of adjuvant chemotherapy in line with national guidelines.

Results: Higher levels of sNRP-1 reflected more advanced disease (FIGO III/IV) and indicated a trend towards suboptimal surgical outcome, i.e. any residual tumor. sNRP-1 was neither related to the patients’ age nor the BRCA1/2 mutational status. Patients with higher sNRP-1 levels at primary diagnosis had a significantly reduced progression-free survival (PFS) (HR = 0.541, 95%CI: 0.304 – 0.963; p = 0.037) and overall survival (OS) (HR = 0.459, 95%CI: 0.225 – 0.936; p = 0.032). Principal component analysis showed that sNRP-1 levels were unrelated to the circulating hepatocyte growth factor (HGF) and the soluble ectodomain of its receptor the tyrosine kinase mesenchymal-epithelial transition (c-MET), suggesting that there is no proportional serological concentration gradient of soluble components of the NRP-1/HGF/c-MET signaling axis.

Conclusions: In line with the previously shown tissue-based prognostic role, we demonstrated for the first time that sNRP-1 can also act as a readily accessible, prognostic biomarker in the circulation of patients with ovarian cancer at primary diagnosis. Given its known role in angiogenesis and conferring resistance to the poly ADP-ribose polymerase (PARP) inhibitor olaparib in vitro, our results encourage more detailed investigation into sNRP-1 as a potential predictive biomarker for bevacizumab and/or PARP-inhibitor treatment.

 

 

 

 

We at BIOMEDICA develop and manufacture ELISA assays to reliably quantify cytokines in biological fluids.

Our ELISA kits are rigorously validated according to international quality standards to ensure their consistency, specificity, precision, and robustness.

All of the validation data can be found on the respective product pages on our website www.bmgrp.com . 

Quantify Cytokines with an ELISA Assay from BIOMEDICA

We designed our assays to quantify the cytokine of interest in serum, plasma or cell culture supernatants.

Standardized harmonization of results with WHO reference reagents

To ensure a harmonized standardization, WHO reference reagents / standards are employed for cross-comparison of results (1)

Quantify Cytokines with an ELISA Assay from BIOMEDICA

IL-6 ELISA (cat. no. BI-IL6)

The IL-6 (Interleukin-6) ELISA kit from BIOMEDICA is a conventional  sandwich ELISA kit. The assay incorporates two epitope-mapped antibodies that specifically bind to human IL-6 in the respective samples.

Capture antibody : is a  recombinant IL-6 antibody that is specific for human IL-6, which is pre-coated on a 96-well microtiter plate. 

Detection antibody: is a polyclonal IL-6 antibody (labeled with streptavidin-HRPO), which is specific for human IL-6.

• • Assay method: sandwich ELISA
  • Sample types: serum, plasma, cell-culture, urine
  • Sample volume: 100µl / well
  • Assay time: 4.5 hrs
  • Standard range: 0; 3.1 – 200 pg/ml
  • Sensitivity: LLOQ: 0.78 pg/ml (measurable concentrations in both serum AND plasma samples!),  LOD: 0.28 pg/ml
  • Precision: in-between-run (n=9 ): ≤ 6 % CV, within-run (n=3): ≤ 7 % CV
  • Specificity:  the ELISA recognizes endogenous (natural) and recombinant and human IL-6
  • Standard matrix: 8 x pre-diluted standards, lyophilized,  in a serum based matrix containing recombinant human Il-6
  • Controls: 1x high and 1x low control included in the kit

 

VEGF ELISA (cat. no. BI-VEGF)

The VEGF (Vascular Endothelial Growth Factor) ELISA kit from BIOMEDICA is a conventional sandwich ELISA kit. The assay incorporates two epitope-mapped antibodies that specifically bind to human VEGF in the respective samples.

Capture antibody is a recombinant VEGF antibody that is specific for human VEGF, which is pre-coated on a 96-well microtiter plate,  recognizing a structural epitope in the conserved receptor binding- site of the VEGF peptide. All VEGF isoforms are detected.

Detection antibody: is a polyclonal VEGF antibody (labeled with streptavidin-HRPO), which is specific for human VEGF. The antibody recognizes multiple linear epitopes that are concentrated in the first 120 amino acids in the N-terminal region of the VEGF molecule.   

• Assay method: sandwich ELISA
• Sample types: serum, plasma, cell-culture, urine
• Sample volume: 10µl / well
• Assay time: 4.5 hrs
• Standard range: 0; 31.2 – 2000 pg/ml
• Sensitivity: LLOQ: 15.6 pg/ml); LOD: 2.5 pg/ml
• Precision: in-between-run: (n=3): ≤ 6 % CV, Within-run (n=3): ≤ 3 % CV
• Specificity: the ELISA recognizes endogenous (natural) human VEGF including all circulating VEGF isoforms including VEGF165b and recombinant VEGF
• Standard matrix: 8 x vials of pre-diluted standards, lyophilized, in a serum based matrix containing recombinant human VEGF
• Controls: 2 controls (high and low) included in the kit

 

ANGIOPOIETIN-2 ELISA (cat. no. BI-ANG2)

The ANG2 (Angiopoietin-2) ELISA kit from BIOMEDICA  is a conventional sandwich ELISA kit. The assay incorporates two epitope-mapped antibodies that specifically bind to human Angiopoietin-2 in the respective samples.

Capture antibody is a recombinant ANG2 antibody that is specific for human VEGF, which is pre-coated on a 96-well microtiter plate. The capture antibody has a structural epitope that binds to the receptor binding site of human Angiopoietin-2. The receptor binds to TEL / TIE2 protein.

Detection antibody: is a polyclonal ANG2 antibody (labeled with streptavidin-HRPO), which is specific for human ANG2. The antibody recognizes several linear epitopes that are distributed over the entire Angiopoietin-2 molecule.  

• Assay method: sandwich ELISA
• Sample types: serum, plasma, cell-culture, urine
• Sample volume: 20µl / well
• Assay time: 3.5 hrs
• Standard range: 0; 0.68 – 21.96 ng/ml (= 0 ; 12.5 – 400 pmol /l  (conversion 1 pmol/l = 54.9 pg/ml (MW: 54.9 kDa)
• Sensitivity: LLOQ : 0.34 ng/ml (=6.3 pmol/l); LOD: 0.20 pg/ml (= 3.7 pmol/l)
• Precision: In-between-run (n=9): ≤ 6 % CV, Within-run (n=3): ≤ 8 % CV
• Specificity: the ELISA recognizes endogenous (natural) human ANG2 and recombinant ANG2.  Most probably all three ANG2 isoforms are detected by the assay, as determined by epitope mapping and analysis of the ANG2 sequence. This ELISA assay does not detect ANG1.
• Standard matrix: 8 x vials of pre-diluted standards, lyophilized, in a serum based matrix containing recombinant human ANG2.
• Controls: 2 controls (high and low) included in the kit

 

Mouse /rat Angiopoietin-2 ELISA (cat. no. BI-ANG2MR) also available

Related publications

1. International reference reagents
2. Angiopoietin-2 Promotes Pathological Angiogenesis and Is a Therapeutic Target in Murine Nonalcoholic Fatty Liver Disease. Lefere S, Van de Velde F, Hoorens A, Raevens S, Van Campenhout S, Vandierendonck A, Neyt S, Vandeghinste B, Vanhove C, Debbaut C, Verhelst X, Van Dorpe J, Van Steenkiste C, Casteleyn C, Lapauw B, Van Vlierberghe H, Geerts A, Devisscher L. Hepatology. 2019 Mar;69(3):1087-1104. doi: 10.1002/hep.30294. Epub 2019 Feb 12. PMID: 30259536.

The development of heart failure is a concerning complication for patients who undergo heart valve and/or coronary bypass grafting surgery. In this context, risk prediction tools for patients undergoing cardiac surgery are of great interest. This article highlights a recent study investigating the predictive power of preoperative FGF23 levels in patients undergoing cardiac surgery.

Fibroblast growth factor 23 (FGF23)

Fibroblast growth factor 23 (FGF23) is a protein that plays a vital role in regulating phosphate and vitamin D levels and is associated with an elevated cardiovascular risk. A recent study by Hofer F et al.,  aimed to explore how FGF23 affects cardiovascular outcomes in a group of patients who underwent cardiac surgery. The analysis included a cohort of 451 patients who were observed for a median duration of 3.9 years. The results indicated that individuals with higher FGF23 levels showed elevated incidence of cardiovascular death. Therefore, the assessment of FGF23 may be a tool for risk stratification and could allow early identification of patients who have a high risk for adverse cardiovascular outcomes.

“Plasma FGF‐23 concentrations were assessed with a sandwich enzyme immunoassay (Biomedica Medizinprodukte GmbH, Vienna, Austria). The intra‐assay and interassay precision values were ≤12% and ≤10%, respectively. The assay limit of detection (0 pmol/L+3 SDs) was 0.08 pmol/L. For better clinical evaluation, FGF‐23 was converted to units of pg/mL, with a conversion factor of 1 pg/mL=0.133 pmol/L.”

Predictive power of preoperative FGF23 levels in patients undergoing cardiac surgery

Routine measurement of preoperative FGF23 levels may improve the detection of high-risk patients undergoing cardiac surgery.

 BIOMEDICA  FGF23 ELISA assays for  serum & plasma samples

FGF23 (c-terminal) ELISA (cat. no BI-20702) and FGF23 intact ELISA (cat. no.  BI-20700)

• developed & manufactured by Biomedica , Vienna, Austria
• fully validated according to international quality guidelines
• numerous citations in top journals

 

Related Publications

Relationship of Fibroblast Growth Factor 23 With Hospitalization for Heart Failure and Cardiovascular Outcomes in Patients Undergoing Cardiac Surgery. Hofer F, Hammer A, Pailer U, Koller L, Kazem N, Steinacher E, Steinlechner B, Andreas M, Laufer G, Wojta J, Zelniker TA, Hengstenberg C, Niessner A, Sulzgruber P. J Am Heart Assoc. 2023 Mar 7;12(5):e027875. doi: 10.1161/JAHA.122.027875. Epub 2023 Feb 21. PMID: 36802737; PMCID: PMC10111457.

Abstract

Background Fibroblast growth factor 23 (FGF-23) is crucial in regulating phosphate and vitamin D metabolism and is moreover associated with an increased cardiovascular risk. The specific objective of this study was to investigate the influence of FGF-23 on cardiovascular outcomes, including hospitalization for heart failure (HHF), postoperative atrial fibrillation, and cardiovascular death, in an unselected patient population after cardiac surgery. Methods and Results Patients undergoing elective coronary artery bypass graft and/or cardiac valve surgery were prospectively enrolled. FGF-23 blood plasma concentrations were assessed before surgery. A composite of cardiovascular death/HHF was chosen as primary end point. A total of 451 patients (median age 70 years; 28.8% female) were included in the present analysis and followed over a median of 3.9 years. Individuals with higher FGF-23 quartiles showed elevated incidence rates of the composite of cardiovascular death/HHF (quartile 1, 7.1%; quartile 2, 8.6%; quartile 3, 15.1%; and quartile 4, 34.3%). After multivariable adjustment, FGF-23 modeled as a continuous variable (adjusted hazard ratio for a 1-unit increase in standardized log-transformed biomarker, 1.82 [95% CI, 1.34-2.46]) as well as using predefined risk groups and quartiles remained independently associated with the risk of cardiovascular death/HHF and the secondary outcomes, including postoperative atrial fibrillation. Reclassification analysis indicated that the addition of FGF-23 to N-terminal pro-B-type natriuretic peptide provides a significant improvement in risk discrimination (net reclassification improvement at the event rate, 0.58 [95% CI, 0.34-0.81]; P<0.001; integrated discrimination increment, 0.03 [95% CI, 0.01-0.05]; P<0.001). Conclusions FGF-23 is an independent predictor of cardiovascular death/HHF and postoperative atrial fibrillation in individuals undergoing cardiac surgery. Considering an individualized risk assessment, routine preoperative FGF-23 evaluation may improve detection of high-risk patients.

A single preoperative FGF23 measurement is a strong predictor of outcome in patients undergoing elective cardiac surgery: a prospective observational study. Speer T, Groesdonk HV, Zapf B, Buescher V, Beyse M, Duerr L, Gewert S, Krauss P, Poppleton A, Wagenpfeil S, Fliser D, Schaefers HJ, Klingele M.Crit Care. 2015 Apr 23;19(1):190. doi: 10.1186/s13054-015-0925-6. PMID: 25902817; PMCID: PMC4424828.

Medication-related osteonecrosis of the jaw (MRONJ) is a severe side effect that can occur in patients taking medications often used for treating cancer and osteoporosis. MRONJ occurs when blood supply to a part of the bone is disrupted. This leads to a gradual destruction of bone in the jaw.

Semaphorin 4D (SEMA4D) is a protein that has various important functions in both the immune- and the nervous system. SEMA4D also plays a role in bone biology as it influences bone remodeling by interacting between immune and bone cells, specifically osteoclasts, the cells responsible for breaking down bone.

SEMAPHORIN 4D in medication-related osteonecrosis of the jaw

In a recent study researchers investigated the clinical significance of circulating SEMA4D in the early diagnosis of MRONJ:

Clinical values of serum Semaphorin 4D (Sema4D) in medication‑related osteonecrosis of the jaw. 

Semaphorin 4D can easily be measured in blood samples with an ELISA assay

Biomedica offers a high-quality test kit to measure soluble SEMA4D in human plasma samples

Semaphorin 4D (SEMA4D) ELISA Assay Highlights:

• • Only Sema4D ELISA assay that is fully validated
  • 10 µl / well sample volume – sample predilution is not required
  • Available reference values for healthy individuals

To see the protocol booklet for SEMA4D ELISA just click here

Related literature

 Clinical values of serum Semaphorin 4D (Sema4D) in medication‑related osteonecrosis of the jaw. Mu H, Pang Y, Liu L, Liu J, Liu C. Eur J Med Res. 2023 Mar 30;28(1):140. doi: 10.1186/s40001-023-01095-6. PMID: 36998031; PMCID: PMC10061851.

Abstract

Background: Bisphosphonates (BPs) are widely used in clinical practice to prevent and treat bone metabolism-related diseases. Medication-related osteonecrosis of the jaw (MRONJ) is one of the major sequelae of BPs use. Early prediction and intervention of MRONJ are of great significance.

Methods: Ninety-seven patients currently on treatment with BPs or with a history of BPs usage and 45 healthy volunteers undergoing dentoalveolar surgery were included in this study. Participants’ serum Semaphorin 4D (Sema4D) levels were measured and analyzed before participants underwent surgery (T0) and after a 12-month follow-up (T1). Kruskal-Wallis test and ROC analysis were used to examine the predictive effect of Sema4D on MRONJ.

Results: Sema4D levels in serum of patients corresponding to confirmed MRONJ were significantly lower at both T0 and T1 time points compared to non-MRONJ and healthy controls. Sema4D has a statistically predictive effect on the occurrence and diagnosis of MRONJ. Serum Sema4D levels were significantly reduced in MRONJ class 3 patients. MRONJ patients who received intravenous BPs had significantly lower Sema4D levels than those who received oral BPs.

Conclusion: Serum Sema4D level has predictive value for the onset of MRONJ in BPs users within 12 weeks after dentoalveolar surgery

Interventions for managing medication-related osteonecrosis of the jaw. Beth-Tasdogan NH, Mayer B, Hussein H, Zolk O. Cochrane Database Syst Rev. 2017 Oct 6;10(10):CD012432. doi: 10.1002/14651858.CD012432.pub2. Update in: Cochrane Database Syst Rev. 2022 Jul 12;7:CD012432. PMID: 28983908; PMCID: PMC6485859.

Possible pathogenic engagement of soluble Semaphorin 4D produced by γδT cells in medication-related osteonecrosis of the jaw (MRONJ). Movila A, Mawardi H, Nishimura K, Kiyama T, Egashira K, Kim JY, Villa A, Sasaki H, Woo SB, Kawai T. Biochem Biophys Res Commun. 2016 Nov 4;480(1):42-47. doi: 10.1016/j.bbrc.2016.10.012. Epub 2016 Oct 5. PMID: 27720716.

 

.

With advancing age, the human body´s natural capacity to consistently renew bones and maintain their resilience diminishes. This decline is enhanced by conditions like osteoporosis. This poses a significant health concern for the elderly and is becoming a growing economic challenge on society. In an effort to address this problem, scientists are actively seeking novel therapeutic strategies to enhance bone regeneration.

 

Building bone with bioinspired molecules

By employing computer models and simulations, a Dresden-based team created innovative bio-inspired compounds that enhance bone regeneration in mice. These compounds can be incorporated into biomaterials, allowing for their localized introduction into bone defects. These newly developed molecules derive from glycosaminoglycans, which are extended sugar chains like hyaluronic acid or heparin. These molecules could be used to turn-off proteins like DKK-1 and Sclerostin that block bone regeneration that could lead to new and more effective therapies for bone diseases.

DKK-1 and SCLEROSTIN are two important proteins that play significant roles in regulating bone health and development.

DKK-1 (Dickkopf-1) acts as an inhibitor of the Wnt signaling pathway. This pathway is crucial for bone formation. Overexpression of DKK-1 leads to a decrease in bone formation characterized by conditions with impaired bone density and increased fragility, such as osteoporosis.

SCLEROSTIN (SOST) is a Wnt signaling inhibitor and a negative regulator of bone formation. Sclerostin is primarily produced by osteocytes, which are bone cells embedded within the bone matrix.

Sclerostin and DKK-1 can be measured in human serum samples with an ELISA assay.

 

Biomedica´s DKK-1 ELISA assay (cat. no. BI-20413)

• 20µl sample volume
• Widely cited
• No sample dilution
• Fully validated

 

Biomedica´s Sclerostin ELISA assay (cat. no. 20492)

• The internationally most referenced Sclerostin ELISA!
• Rigorously validated according to FDA/ICH/EMEA guidelines
• 20µl sample volume

 

also available: Bioactive Sclerostin ELISA (cat. no. BI-20472)

Rational engineering of glycosaminoglycan-based Dickkopf-1 scavengers to improve bone regeneration. Ruiz-Gómez G, Salbach-Hirsch J, Dürig JN, Köhler L, Balamurugan K, Rother S, Heidig SL, Moeller S, Schnabelrauch M, Furesi G, Pählig S, Guillem-Gloria PM, Hofbauer C, Hintze V, Pisabarro MT, Rademann J, Hofbauer LC. Biomaterials. 2023 Jun;297:122105. doi: 10.1016/j.biomaterials.2023.122105. Epub 2023 Mar 31. PMID: 37031548.

Abstract

The WNT signaling pathway is a central regulator of bone development and regeneration. Functional alterations of WNT ligands and inhibitors are associated with a variety of bone diseases that affect bone fragility and result in a high medical and socioeconomic burden. Hence, this cellular pathway has emerged as a novel target for bone-protective therapies, e.g. in osteoporosis. Here, we investigated glycosaminoglycan (GAG) recognition by Dickkopf-1 (DKK1), a potent endogenous WNT inhibitor, and the underlying functional implications in order to develop WNT signaling regulators. In a multidisciplinary approach we applied in silico structure-based de novo design strategies and molecular dynamics simulations combined with synthetic chemistry and surface plasmon resonance spectroscopy to Rationally Engineer oligomeric Glycosaminoglycan derivatives (_REGAG) with improved neutralizing properties for DKK1. In vitro and in vivo assays show that the GAG modification to obtain _REGAG translated into increased WNT pathway activity and improved bone regeneration in a mouse calvaria defect model with critical size bone lesions. Importantly, the developed _REGAG outperformed polymeric high-sulfated hyaluronan (sHA3) in enhancing bone healing up to 50% due to their improved DKK1 binding properties. Thus, rationally engineered GAG variants may represent an innovative strategy to develop novel therapeutic approaches for regenerative medicine

ELISA – Luminex – NGS – microRNA

Analytical Service Measurements – we offer tailored analytical testing services for

ELISA  & Luminex Assays, NGS & microRNA  and can support your research with:

 

• Expertise: experienced laboratory staff
• Quality: highest quality equipment
• Flexibility: tailored solutions according to your project needs and budget
• Speed: rapid turn-around time to meet your deadlines
• Results: verified and comprehensive results presented in an analytical report

Learn more about our ELISA and Luminex Workflow chart and our microRNA service details click here

Analytical Service Measurements – Biomedica

ELISA

We will be happy to measure any kind of biomarker of your choice with an ELISA assay (enzyme linked immunosorbent assay) including our propriety Biomedica biomarker ELISA kits (www.bmgrp.com).

Our Service Measurement Applications include analysis of samples for:

• • Clinical studies (serum and plasma samples)
  • Pre-clinical studies (any species)

 

Of note: Biomedica offers several biomarker ELISA kits for the application in various animal models: Rat-NT-proBNP ELISA and  NT-proANP ELISA (widely used in rat and mice,  works also in also in rabbit or pig models).

• • Custom applications – Learn about our workflow

RNA Services

Together with TamiRNA we offer a range of quality RNA services:

• RNA extraction: Biofluids including serum, plasma, and extracellular vesicles. Cells and tissues – the quality control of total RNA is carried out utilizing bioanalyser chips. 
• Next-generation sequencing- NGS.
• RT-qPCR
• We also analyse cell-type specific microRNA/mRNA  in complex tissues and offer custom analysis of microRNA signatures.
• Finally, we carry out the analysis of established microRNA signatures to predict fracture risk, platelet function, and toxicity.

 

Further reading

Enzyme-Linked Immunosorbent Assay: Types and Applications. Hayrapetyan H, Tran T, Tellez-Corrales E, Madiraju C. Methods Mol Biol. 2023;2612:1-17. doi: 10.1007/978-1-0716-2903-1_1. PMID: 36795355.

RNA Extraction from Cartilage: Issues, Methods, Tips. Pagani S, Maglio M, Sicuro L, Fini M, Giavaresi G, Brogini S. Int J Mol Sci. 2023 Jan 20;24(3):2120. doi: 10.3390/ijms24032120. PMID: 36768444; PMCID: PMC9917073.

A Comprehensive Review of Performance of Next-Generation Sequencing Platforms. Pervez MT, Hasnain MJU, Abbas SH, Moustafa MF, Aslam N, Shah SSM. Biomed Res Int. 2022 Sep 29;2022:3457806. doi: 10.1155/2022/3457806. PMID: 36212714; PMCID: PMC9537002.

Individuals suffering from end-stage kidney disease (ESKD) undergoing dialysis have significantly higher mortality rates compared to patients with non-ESKD of the same age. Approaches to assess mortality risk, which could enable tailored and effective care are needed (1).

Angiopoietin-2 (Ang2) is a protein that is crucial in pathological vascular restructuring and in the formation of new blood vessels (angiogenesis). These processes are active in ESKD patients and might contribute to the elevated mortality observed in this patient group.

Angiopoietin-2 predicts mortality in male ESKD patients on hemodialysis

In a recent multicenter prospective study Ang2 was measured in 340 hemodialysis (HD) patients with ESKD to predict overall mortality (2, 3). The patients were followed up for 5-years and blood samples and clinical data were collected at baseline. Ang2 serum levels were measured with a enzyme-linked immunosorbent assay (ELISA) from Biomedica.

Angiopoietin-2 predicts mortality in male ESKD patients on hemodialysis

The results of the study demonstrated that the risk of death was higher in male patients with elevated Ang2 levels than in female patients: Ang2 levels at baseline are independently associated with all cause mortality in male ESKR patients on HD. 

Angiopoetin-2 can reliably be measured in serum or plasma samples with a conventional ELISA assay: 

Human ANGIOPOIETIN-2 ELISA (cat. no. BI-ANG2) 

• 20µl sample volume /well
• optimized for clinical samples – sample values provided
• kit includes ready to use standards and controls  
• developed & manufactured by BIOMEDICA
• for serum/plasma/cell culture samples
• validation package available

 

also available: 

Mouse / rat ANGIOPOIETIN-2 ELISA  (cat. no. BI-ANG2MR)

• 5µl sample volume /well
• kit control included
• sample values provided

 

Related Publications

1. One- and 2-Year Mortality Prediction for Patients Starting Chronic Dialysis. Haapio M, Helve J, Grönhagen-Riska C, Finne P. Kidney Int Rep. 2017 Jun 24;2(6):1176-1185. doi: 10.1016/j.ekir.2017.06.019. PMID: 29270526; PMCID: PMC5733880.
2. Angiopoietin-2 predicts all-cause mortality in male but not female end-stage kidney disease patients on hemodialysis. Chu C, Chen X, Hasan AA, Szakallova A, Krämer BK, Tepel M, Hocher B. Nephrol Dial Transplant. 2022 Jun 23;37(7):1348-1356. doi: 10.1093/ndt/gfab332. PMID: 34792167; PMCID: PMC9217660.
3. Circulating angiopoietin-2 level is independently associated with all-cause mortality in male end-stage kidney disease patients on hemodialysis. Chu, Chang. Thesis, 2023-06-22T11:52:24Z.

 

 

Lyme disease, also known as Lyme borreliosis, is a tick-borne infectious disease mainly caused by the bacterium species Borrelia burgdorferi.

The disease is primarily transmitted to humans through tick bites. Skin rash, fever, headache, fatigue, and joint pain are among the symptoms characterized by the disease. If left untreated, the infection can lead to more severe conditions and can spread to various parts of the body, affecting joint, heart, and nervous system. Lyme disease is most prevalent in temperate regions, particularly in North America, Europe, and parts of Asia. The incidence of Lyme disease has been increasing in recent years, making it an important health concern.

Early diagnosis and treatment are essential for managing Lyme disease effectively.

Testing for Lyme Borreliosis

The BIOMEDICA Borrelia ELISA kits were utilized in a recent study analyzing antibody patterns that could be useful in guiding the diagnostic schedule: Testing for Lyme borreliosis: could serology tell more? Zóka A et al., J Immunol Clin Microbiol. 2020; 5(3): 85-93. link .

Borrelia IgG ELISA | BI-21032

Recombinant Antigens– therefore:

• High sensitivity and specificity confirmed by clinical samples
• Standardized CSF and serum analysis available
• For manual and automated testing
• Widely cited

 

Lyme borreliosis is a bacterial infection caused by the spirochaete Borrelia burgdorferi, afzellii or garinii, and is characterised by a variety of clinical symptoms.

 Lyme borreliosis can be divided into 3 stages:

 Stage 1, early dermatitis, Clinical: erythema migrans.

Stage 2, early disseminated infection, Clinical: lymphocytic meningoradiculitis (Bannwarth’s syndrome), neuroborreliosis.

Stage 3, late disseminated infection, Clinical: chronic progressive encephalomyelitis, acrodermatitis chronica atrophicans (ACA), chronic arthritis.

To improve the diagnostic specifity, the Biomedica Borrelia IgG ELISA microwell strips are coated with the following recombinant antigens:

• p21 = OspC (outer surface protein C): B. burgdorferi sensu stricto (B31), B. garinii (20047)
• p18: B. afzelii (pKo)
• p100: B. afzelii (pKo)
• VIsE: fusion protein of different Borrelia genospecies

 

Borrelia IgM ELISA | BI-21042

Recombinant Antigens– therefore:

• No extra RF stripping necessary
• High sensitivity and specificity confirmed by clinical samples
• Standardized CSF-serum-analysis available
• For manual and automated testing
• Widely cited

 

To improve the diagnostic specifity, the Biomedica Borrelia IgM ELISA microwell strips are coated with the following recombinant antigens:

• p21 = OspC (outer surface protein C): B. afzellii (pKo)
• p21 = OspC (outer surface protein C): B. garinii (20047)
• p41/I = (inner part of flagellin): B. bavariensis (pBi)
• VIsE: fusion proteins of different Borrelia genospecies 

 

Related publications

Lyme borreliosis diagnosis: state of the art of improvements and innovations. Guérin M, Shawky M, Zedan A, Octave S, Avalle B, Maffucci I, Padiolleau-Lefèvre S. BMC Microbiol. 2023 Aug 1;23(1):204. doi: 10.1186/s12866-023-02935-5. PMID: 37528399; PMCID: PMC10392007.

The Epidemiology of Lyme Borreliosis in Europe: An Updated Review on a Growing Public Health Issue. Stark JH, Pilz A, Jodar L, Moïsi JC.Vector Borne Zoonotic Dis. 2023 Apr;23(4):139-141. doi: 10.1089/vbz.2022.0068. Epub 2023 Jan 27. PMID: 37071398; PMCID: PMC10122224.

Effectiveness of personal protection measures against Lyme disease: A review of epidemiologic studies from the United States. Schwartz AM, Mackeprang JM, Mead PS, Hinckley AF. Zoonoses Public Health. 2022 Nov;69(7):777-791. doi: 10.1111/zph.12984. Epub 2022 Jul 5. PMID: 35791092.

Breast cancer is the most common cancer in women worldwide. Approximately 15-20% of breast cancer cases are Human Epidermal Growth Factor Receptor (HER2)-positive. The overexpression of the HER2 protein promotes the growth and spread of cancer cells, indicating a poorer prognosis in these patients. Therapies, known as anti-HER2 drugs, specifically target the HER2 protein and inhibit its activity, thus improving the survival of patients with HER2 positive breast cancer. 

Evaluating Cardiotoxicity of HER-2 Targeted Drugs for Breast Cancer

One known side effect of HER-2 targeted therapies is cardiotoxicity which is characterized by left ventricular systolic dysfunction. Identifying cardiac dysfunction in HER2 positive breast cancer patients is crucial in order to prevent the development of heart failure in these patients. Monitoring cardiac function through imaging methods is mandatory for HER2-positive breast cancer patients. However, considering the possibility of cardiac damage even if there are no evident changes in heart function could be beneficial. Therefore, biomarkers could be useful in assessing early cardiac dysfunction in these patients.

Evaluating Cardiotoxicity of HER-2 Targeted Drugs for Breast Cancer: 

The cardiac biomarkers  NT-proBNP and NT-proANP

N-terminal pro-brain natriuretic peptide (NT-proBNP – amino acids 1-76) and N-terminal pro-atrial natriuretic peptide (NT-proANP – amino acids 1-98) are secreted from the cardiac ventricles and atria, respectively. The half-lives and stability of both analytes are more reliable than their biologically active peptides (BNP and ANP).

In a recent study researchers evaluated patients diagnosed with HER-2 positive breast cancer who underwent treatment with HER2 inhibitors and measured serum levels of the cardiac biomarkers NT-proBNP and NT-proANP using the assays from Biomedica. Learn more:

Evaluating Cardiotoxicity in Breast Cancer Patients Treated with HER2 Inhibitors: Could a Combination of Radionuclide Ventriculography and Cardiac Biomarkers Predict the Cardiac Impact?

Ready tp use kits developed & manufactured by BIOMEDICA

 

NT-proBNP ELISA Assay (cat. no SK-1204)

•  For the detection of human NT-proBNP
• CE.marked – for IVD use in the EU
• Flexible – can be run in every lab
• 20µl sample volume (serum/plasma)
• Assay time – 3 h / 30 min
• Two controls and ready to use calibrators included
• Regular proficiency testing –“Proficiency Testing Certificate”

 

NT-proANP ELISA Assay (cat. no BI-20892)

• For the detection of human NT-proANP (assay suitable also for detection of NT-proANP in rat and mouse samples)
• Two controls and ready to use calibrators included
• 50µl sample volume (serum/plasma)
• Assay time – 3 h / 30 min

 

Related products

Rat NT-proBNP ELISA assay (cat. no. BI-1204R)

• 10 µl sample volume
• sample values provided

 

Related literature

Evaluating Cardiotoxicity in Breast Cancer Patients Treated with HER2 Inhibitors: Could a Combination of Radionuclide Ventriculography and Cardiac Biomarkers Predict the Cardiac Impact? Gherghe M, Lazar AM, Mutuleanu MD, Bordea CI, Ionescu S, Mihaila RI, Petroiu C, Stanciu AE. Cancers (Basel). 2022 Dec 29;15(1):207. doi: 10.3390/cancers15010207. PMID: 36612202; PMCID: PMC9818586.

Abstract

(1) Background: The aim of our study was to determine whether monitoring cardiac function through RNV and cardiac biomarkers could predict the cardiac impact of combined therapy with trastuzumab, pertuzumab and docetaxel, which are regularly used nowadays to treat HER2-positive breast cancer. (2) Methods: This prospective monocentric study included 22 patients, diagnosed with HER2-positive breast cancer, who had their LVEFs and cardiac biomarkers evaluated both at the beginning of their treatment and after 6 months. Among all of the enrolled patients, two blood specimens were collected to assess circulating cardiac biomarkers. RNV was performed in each patient after “in vivo” radiolabeling of the erythrocytes. The obtained results were then statistically correlated. (3) Results: The average LVEF decrease between the two time points was approximately 4%. Of the five biomarkers we considered in this paper, only NT-proBNP correlated with the LVEF values obtained both in the baseline study and after 6 months of follow-up (r = -0.615 for T0 and r = -0.751 for T1, respectively). ST2/IL-33R proved statistically significant at the T1 time point (r = -0.547). (4) Conclusions: A combination of LVEF, NT-proBNP and ST2/IL-33R assessment may be useful for early detection of cardiac impairment in breast cancer patients treated with trastuzumab, pertuzumab and docetaxel.

NT-proBNP correlates with LVEF decline in HER2-positive breast cancer patients treated with trastuzumab. Bouwer NI, Liesting C, Kofflard MJM, Sprangers-van Campen SM, Brugts JJ, Kitzen JJEM, Fouraux MA, Levin MD, Boersma E. Cardiooncology. 2019 May 28;5:4. doi: 10.1186/s40959-019-0039-4. PMID: 32154011; PMCID: PMC7048136.

NT-proBNP as predictor factor of cardiotoxicity during trastuzumab treatment in breast cancer patients. Blancas I, Martín-Pérez FJ, Garrido JM, Rodríguez-Serrano F. Breast. 2020 Dec;54:106-113. doi: 10.1016/j.breast.2020.09.001. Epub 2020 Sep 11. PMID: 32977298; PMCID: PMC7511727.

We are scientists, developers and manufacturers and understand the importance of offering assays that generate specific, reliable and reproducible results.

Early on, starting from product development to final assay validation and product release until final ELISA kit manufacturing, every assay goes through a stringent quality control process.

Why choose ELISA assays from Biomedica?

Because we care!

Get the most accurate results from your precious samples with Biomedica ELISA kits.

   

 

SPECIFICIC – RELIABLE – REPRODUCIBLE ELISA Assays

 1. SPECIFICITY – epitope-mapped and characterized antibodies for accurate biomarker detection.

The performance of an ELISA is linked to the quality of the antibody pairs used for biomarker detection.

We therefore:

• select antibody pairs with high affinity and specificity with mapped binding sites
• optimize our assays to quantify biomarkers in both healthy and pathological samples

 

Example: FGF23 sample values of normal and pathological samples

2. RELIABILITY – extensive validation using clinical samples (parallelism, S/R, precision, analyte stability..) in various sample matrices

We validate our ELISA assays according to international quality guidelines (FDA, EMEA) .

Our Biomedica Immunassays go through an extensive validation process 

2.1. Accuracy – accurate detection of biomarkers in clinical samples and exclusion of matrix effects that may interfere with the measurement of the analyte of interest.

• Accuracy is determined in all validated sample types that are spiked with known amounts of the recombinant analyte. Samples are  analysed against the standard/calibration curve of the assay and then compared with the nominal value.

 

2.2. Parallelism / Dilution Linearity – lot to lot consistency ensured by our stringent quality management guidelines

• During assay validation we analyze the recovery of the analyte in diluted samples that contains the endogenous / recombinant analyte of interest.

 

Example: dilution linearity (parallelism) of samples containing endogenous and recombinant Neuropilin-1 (NRP1).

2.3. Specificity and Cross-Reactivity – only detects the analyte of interest

• We carefully select antibodies that exclusively detect the specific analyte. Our specificity experiments are designed to characterize the antibody-antigen interactions and to determine possible isoforms that could be bound by the antibody.

 

Example: antibody recognizing all three isoforms of the target analyte on a Western blot.

 

2.4. Sensitivity

• Our ELISA assays are optimized to minimize the background signal while maximizing the signals from the measurements of the analyte ensuring maximal sensitivity.

The data for the Lower Limit of Quantification (LLOQ) and the Limit of Detection (LOD) are indicted in the instructions for use and on our website for all our ELISA kits.

2.5. Precision – precise and reproducible results within and across lots

• Within-run and in-between run precision is tested several times within one ELISA assay lot to guarantee that results are accurate when using kits that derive from different lots.

2.6. Calibration

• The accurate quantification of a biomarker depends on the linearity and the reproducibility of the standard curve. During the assays optimization process we ensure low variability between the results of the calibrators. Where available, we employ WHO reference reagents to ensure a harmonized standardization.

 

Example: standard curve for the FGF23 ELISA after 4PL transformation. The error bars reflect the variability of the measurement.

2.7. Stability

• During development we test the stability of all assay components as well as the stability of the analyte of interest in the respective sample matrices (serum, plasma). For instance we expose real clinical samples with elevated levels of the analyte to multiple freeze-thaw cyles and also determine stability at room temperature.

 

Example: stability of the analyte Periostin in clinical samples after multiple freeze-thaw (F/T) cycles in different sample matrices.

 

Freeze-thaw stability of Periostin

 

Validation reports

The validation reports of the respective ELISA assays can be  downloaded on the individual Biomedica ELISA product pages.

 

3. REPRODUCIBILITY

 ELISA ASSAY QUALITY MANAGEMENT

 Our Quality Control Process

All our kits undergo a stringent quality control process, including testing of lot-to-lot consistency as well as the kit stability during shelf-life.

Our manufacturing process follows the ISO 9001: 2015 management system and conforms to GMP /GLP guidelines.

Ensuring lot-to-lot consistency with a panel of quality control samples

Our internal quality control panel is one integral part of our manufacturing protocols. It contains samples from different matrices (serum, EDTA-plasma, citrate-plasma..) containing the endogenous/natural analyte as well as samples spiked with the recombinant protein. Every new lot as well as all retains, that are assayed every three months, are tested with the specific QC sample panel.

Example:  IC trending showing the quotient of the proANP concentration measured in Internal Controls (IC) in 3 different proANP ELISA lots compared with previously established median concentrations.

 

 

 

Qualified CUSTOMER SERVICE – we accompany you in every step.

Our qualified customer service representatives have hands-on research experience to assist you along the way, from decision making to technical questions.

WE VALUE YOUR OPINION

Our ELISA assays are developed to serve your needs. We therefore select our biomarker targets based on your input. 

Pre-testing: before a new ELISA assay is launched,  selected customers test the product to ensure that the assay is reliable and reproducible outside of our lab-facilities. 

Biomedica – ELISA development scheme  

 

Learn more about how we guarantee the performance of our products – click here .

 

Further reading

ICH Q2(R2) Validation of analytical procedures – Scientific guideline