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• Why choose ELISA assays from Biomedica?

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

 

Major depressive disorder (MDD) is a prevalent mental health condition that ranks among the top psychiatric illnesses worldwide. Approximately 30-60% of patients diagnosed with depression do not show positive responses to currently available antidepressant treatments. There is consistent evidence suggesting that elevated blood levels of the cytokine interleukin-6 (IL-6) in individuals with MDD have a significant impact on stress responses and correlate with depression severity scores (1, 2).

IL-6 in depressive disorder

Given the association between IL-6 and MDD, inhibiting IL-6 has gained attention as a potential therapeutic approach for depression. By targeting IL-6 pathways it is hypothesized that the aberrant inflammatory response associated with MDD can be modulated, leading to improved treatment outcomes. Exploring IL-6 inhibition, as a novel strategy holds promise for the development of more effective therapies for individuals who are non-responsive to current antidepressant treatments.

IL-6 can reliably be measured in various sample types by ELISA assay:

BIOMEDICA IL-6 ELISA (cat. no. BI-IL6)

• Sensitive – Measurable values in serum and plasma samples
• Reliable – Full validation package
• Specific-  Characterized epitope-mapped antibodies
• Easy – Color-coded reagents and controls included

 

Related ELISA products

VEGF (cat. no. BI-VEGF), Angiopoietin-2 (BI-ANG2), Big-Endothelin (cat. no. BI-20082H), NT-proCNP (cat. no. BI-20812)

Publications

1. How does IL-6 change after combined treatment in MDD patients? A systematic review. Lombardi AL, Manfredi L, Conversi D. Brain Behav Immun Health. 2022 Dec 24;27:100579. doi: 10.1016/j.bbih.2022.100579. PMID: 36624849; PMCID: PMC9822965.
2. Role of Interleukin-6 in Depressive Disorder. Ting EY et al., Int J Mol Sci. 2020 Mar 22;21(6):2194. doi: 10.3390/ijms21062194. PMID: 32235786; PMCID: PMC7139933.

 

Related literature

Interleukin 6 as a marker of depression in women with sleep apnea. Campos-Rodriguez F et al., J Sleep Res. 2021 Feb;30(1):e13035. doi: 10.1111/jsr.13035. Epub 2020 Mar 25. PMID: 32212220.

 

Abstract

Depression is common in women with obstructive sleep apnea (OSA), but objective markers of depression have not yet been explored in such patients. We hypothesized that inflammation and antioxidant biomarkers may be associated with depression in a cohort of OSA women. We conducted a multicentre, cross-sectional study in 247 women diagnosed with moderate-to-severe OSA. Depression was assessed by the depression subscale of the Hospital Anxiety and Depression Questionnaire (HAD-D) and defined as a score ≥11. Associations between tumour necrosis factor α (TNFα), interleukin 6 (IL-6), C-reactive protein (CRP), intercellular adhesion molecule 1 (ICAM-1), catalase (CAT), superoxide dismutase (SOD) and brain-derived neurotrophic factor (BDNF) plasma levels and depression were assessed. The women had a median (25th-75th percentiles) age of 58 (51-65) years, body mass index (BMI) of 33.5 (29.0-38.3) Kg/m² , Epworth Sleepiness Scale (ESS) score of 10 (6-13) and apnea-hypopnea index (AHI) of 33.3 (22.8-49.3). Logistic regression analyses revealed that only IL6 levels were associated with the presence of depression (adjusted odds ratio [OR], 1.20; 95% confidence interval [CI], 1.08-1.34), whereas linear regression further confirmed that IL6 levels were significantly associated with HAD-D scores (β = .154; 95% CI, 0.03-0.30). Multivariate regression analysis showed that IL6 (OR, 1.22; 95% CI, 1.09-1.36), ESS (OR, 1.10; 95% CI 1.02-1.19) and physical activity <30 min/day (OR, 2.51; 95% CI, 1.25-5.05) were independent predictors of depression. Thus, we conclude that in a cohort of women with moderate-to-severe OSA, IL6 levels are independently associated with the presence of depression and correlate with depression scores. Low physical activity and higher ESS scores are also independent indicators of risk of depression in this population.

The process of aging is linked to physiological changes, which include a decline in bone mass and renal function. In a recent study, researchers investigated the effects of Vitamin D supplementation on bone metabolism and bone biomarkers in patients with and without renal impairment.

Effects of Vitamin D on Bone Markers in Kidney Disease

The study incorporated 379 patients with a mean age of > 70 years who were supplemented with various doses of vitamin D ranging from 12000 IU to 48,000 IU/month for a period of one year.

The biomarkers Sclerostin (SOST), Dickkopf-1 (DKK-1), Osteoprotegerin (OPG), and soluble RANKL (sRANKL) were measured with ELISA assays from BIOMEDICA.

Learn more: Early renal impairment affects hormonal regulators of calcium and bone metabolism and Wnt signalling and the response to vitamin D supplementation in healthy older adults. Christodoulou M et al., J Steroid Biochem Mol Biol. 2023. 229:106267. doi: 10.1016/j.jsbmb.2023.106267. 

About the BIOMEDICA bone marker ELISA kits

Sclerostin (SOST) ELISA  (BI-20492)

• Most referenced Sclerostin ELISA + 250 citations
• Low sample volume – 20µl / well
• Validated following international guidelines

 

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 +245 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 + 290 publications

 

FGF23 intact ELISA (BI-20700)

• for serum and plasma samples
• full validation data available
• citations

 

FGF23 c-terminal ELISA (BI-20702)

• for serum and plasma
• full validation data available
• + 43 citations

 

All assays are developed and manufactured by BIOMEDICA

Related publications

Fibroblast growth factor 23 (FGF23) and early chronic kidney disease in the elderly. Chudek J et al., Nephrol Dial Transplant. 2014 Sep;29(9):1757-63. doi: 10.1093/ndt/gfu063. 

Effectiveness and safety of vitamin D in relation to bone health. Cranney A et al., Evid Rep Technol Assess (Full Rep). 2007 Aug;(158):1-235. PMID: 18088161; PMCID: PMC4781354.

Bone markers are currently used to monitor skeletal diseases and treatments. The proteins Sclerostin and Dickkopf-1 (DKK-1) reflect distinct biological processes and have gained attention as potential biomarkers for bone-related conditions. They may provide valuable information for diagnosis, prognosis, and monitoring of bone diseases and treatments.

Sclerostin and DKK-1 emerging biomarkers for bone disease

Sclerostin and Dickkkopf-1 are two important osteocyte proteins that are involved in the regulation of bone metabolism, particularly through their interactions with the Wnt signaling pathway.

SCLEROSTIN (SOST) is a glycoprotein that is primarily secreted by osteocytes, the most abundant cells in bone tissue. It inhibits Wnt signaling, which is a critical pathway regulating bone formation and remodeling. Sclerostin acts as a negative regulator of bone formation by binding to the LRP5/6 co-receptors (low-density lipoprotein receptor protein), which activate Wnt signaling. By binding to LRP5/6, sclerostin inhibits the interaction between Wnt ligands and the Frizzled receptor, thereby inhibiting Wnt signaling and suppressing bone formation. Inhibition of Sclerostin has led to the development of a novel anabolic therapy for osteoporosis.

DICKKOPF-1 (DKK-1) is a protein that also inhibits the Wnt signaling pathway. DKK-1 binds to the LRP5/6 co-receptors thereby preventing Wnt ligand interaction thus inhibiting bone formation and promoting bone resorption.

The Wnt-signaling pathway is one of the most important pathways controlling bone metabolism. Sclerostin and Dickkopf-1 act as Wnt inhibitors and play a crucial role in controlling bone formation and resorption.

Sclerostin and DKK-1 can easily be measured in blood samples with an ELISA assay

BIOMEDICA offers two kits to measure Sclerostin

Bioactive Sclerostin ELISA cat. no. BI-20472

• serum, plasma, cell-culture
• characterized antibodies
• day test
• 20µl sample volume
• sample values provided
• full validation package

 

The epitope of the monoclonal capture antibody is in loop 2 of the Sclerostin molecule, which is the binding site for the LRP5/6 complex.

All Sclerostin molecules including potential fragments containing this receptor binding region can be detected.

Bioactive Sclerostin ELISA – antibody binding sites

 

Sclerostin ELISA cat. no. BI-20492

• most referenced more than 280 citations
• 20µl sample volume
• for serum, plasma
• full validation package

 

DKK-1 ELISA cat. no. BI-20413

• widely cited
• day test
• no sample dilution
• full validation package

 

All assays are developed and manufactured by BIOMEDICA

Related Publications

New Emerging Biomarkers for Bone Disease: Sclerostin and Dickkopf-1 (DKK1). Dincel AS, Jørgensen NR; IOF-IFCC Joint Committee on Bone Metabolism (C-BM). Calcif Tissue Int. 2023 Feb;112(2):243-257. doi: 10.1007/s00223-022-01020-9. Epub 2022 Sep 27. PMID: 36165920.

Abstract

A healthy skeleton depends on a continuous renewal and maintenance of the bone tissue. The process of bone remodeling is highly controlled and consists of a fine-tuned balance between bone formation and bone resorption. Biochemical markers of bone turnover are already in use for monitoring diseases and treatment involving the skeletal system, but novel biomarkers reflecting specific biological processes in bone and interacting tissues may prove useful for diagnostic, prognostic, and monitoring purposes. The Wnt-signaling pathway is one of the most important pathways controlling bone metabolism and consequently the action of inhibitors of the pathway such as sclerostin and Dickkopf-related protein 1 (DKK1) have crucial roles in controlling bone formation and resorption. Thus, they might be potential markers for clinical use as they reflect a number of physiological and pathophysiological events in bone and in the cross-talk with other tissues in the human body. This review focuses on the clinical utility of measurements of circulating sclerostin and DKK1 levels based on preanalytical and analytical considerations and on evidence obtained from published clinical studies. While accumulating evidence points to clear associations with a number of disease states for the two markers, and thus, the potential for especially sclerostin as a biochemical marker that may be used clinically, the lack of standardization or harmonization of the assays still hampers the clinical utility of the markers.

 

Therapeutic approaches to activate the canonical Wnt pathway for bone regeneration. Nelson AL, Fontana G, Miclau E, Rongstad M, Murphy W, Huard J, Ehrhart N, Bahney C. J Tissue Eng Regen Med. 2022 Nov;16(11):961-976. doi: 10.1002/term.3349. Epub 2022 Sep 16. PMID: 36112528; PMCID: PMC9826348.

The BIOMEDICA FGF23 ELISA assays were recently used in a study investigating hepcidin and iron status in patients with inflammatory bowel disease undergoing therapy (1). Hepcidin, a peptide produced by liver cells, plays a vital role in regulating the body´s iron levels. In instances of inflammation, circulating hepcidin levels rise. Inflammation and iron deficiency have also been shown to stimulate FGF23 production (2). Iron deficiency enhances both the transcription and post-translational cleavage of FGF23 (3). Typically, this results in increased serum FGF23 C-terminal levels, with little to no effect on the biologically active intact FGF23 concentrations (4).

About FGF23

Fibroblast growth factor 23 (FGF23) is a protein that plays a crucial role in regulating phosphate and vitamin D metabolism in the body.

It is primarily produced by bone cells in the tissue called osteocytes. FGF 23 acts on the kidneys and decreases phosphate reabsorption thereby preventing the excessive accumulation of phosphate in the body.

Inflammatory bowel disease and FGF23

Patients with inflammatory bowel disease (IBD) face a greater risk to develop osteopenia and osteoporosis compared to the general population (5). In a study investigating the role of FGF23 in childhood inflammatory bowel disease, serum FGF23 was significantly higher in patients with IBD compared to controls (6)

How can you quantify circulating FGF23 levels?

FGF23 can easily be measured in blood samples (serum or plasma) with a conventional ELISA assay. The levels of FGF23 in the bloodstream consist of both the active intact hormone (iFGF23) and the inactive c-terminal fragments (cFGF23).

The C-terminal FGF23 assay captures both the intact FGF23 hormone and the fragments that form after FGF23 has been cleaved. The intact FGF23 assays utilize antibodies that are positioned in the N-terminal and C-terminal regions of the hormone, specifically targeting the biologically active intact FGF23.

Read more :  FGF23 an Overview

BIOMEDICA  FGF23 ELISA kits for  serum & plasma samples

FGF23 intact ELISA, # BI-20700

FGF23 (c-terminal) ELISA, # BI-20702

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

 

Related products : INTERLEUKIN-6  (IL-6) ELISA , #BI-IL6

• highly sensitive-measurable values in both serum and plasma
• ready to use calibrators and controls included

 

Literature

1. Hepcidin and Iron Status in Patients With Inflammatory Bowel Disease Undergoing Induction Therapy With Vedolizumab or Infliximab. Loveikyte R, Bourgonje AR, van der Reijden JJ, Bulthuis MLC, Hawinkels LJAC, Visschedijk MC, Festen EAM, van Dullemen HM, Weersma RK, van Goor H, van der Meulen-de Jong AE, Dijkstra G. Inflamm Bowel Dis. 2023 Feb 7:izad010. doi: 10.1093/ibd/izad010. Epub ahead of print. PMID: 36748574.
2. Inflammation and functional iron deficiency regulate fibroblast growth factor 23 production. David V, Martin A, Isakova T, Spaulding C, Qi L, Ramirez V, Zumbrennen-Bullough KB, Sun CC, Lin HY, Babitt JL, Wolf M. Kidney Int. 2016 Jan;89(1):135-46. doi: 10.1038/ki.2015.290. Epub 2016 Jan 4. PMID: 26535997; PMCID: PMC4854810.
3. Regulation of Fibroblast Growth Factor 23 by Iron, EPO, and HIF. Wheeler JA, Clinkenbeard EL. Curr Mol Biol Rep. 2019 Mar;5(1):8-17. doi: 10.1007/s40610-019-0110-9. Epub 2019 Jan 25. PMID: 31218207; PMCID: PMC6582956.
4. Regulation of FGF23: Beyond Bone. Simic P, Babitt JL.Curr Osteoporos Rep. 2021 Dec;19(6):563-573. doi: 10.1007/s11914-021-00703-w. Epub 2021 Nov 10. PMID: 34757587; PMCID: PMC8958553.
5. Advances in the understanding of mineral and bone metabolism in inflammatory bowel diseases. Ghishan FK, Kiela PR. Am J Physiol Gastrointest Liver Physiol. 2011 Feb;300(2):G191-201. doi: 10.1152/ajpgi.00496.2010. Epub 2010 Nov 18. PMID: 21088237; PMCID: PMC3043650.
6. Fibroblast growth factor 23 contributes to diminished bone mineral density in childhood inflammatory bowel disease. El-Hodhod MA, Hamdy AM, Abbas AA, Moftah SG, Ramadan AA. BMC Gastroenterol. 2012 May 2;12:44. doi: 10.1186/1471-230X-12-44. PMID: 22551310; PMCID: PMC3438067.

Biomedica features FGF23 ELISA – intact and C-terminal – assays

Meet us at the OSTEOLOGIE Conference taking place from 22-24 June 2023 in Salzburg, Austria! We will exhibit at booth # 36 introducing our novel biomarker ELISA assays to investigate bone and cancer induced bone diseases (e.g. Periostin, Semaphorin 4D, LRG, and others).

Our focus will also be on FGF23, an important regulator of renal phosphate handling, with our FGF23 intact and FGF23 C-terminal ELISA assays.

Drop by – we’d love to see you there!

Related products:

DKK-1 ELISA
OPG ELISA
free sRANKL ELISA
osteomiR – miRNA bone biomarker
FluoBolt Klotho FIA
Testing Services (ELISA, Luminex, miRNA)

 

Rat NT-proBNP and NT-proANP ELISA assays for drug discovery and translational research

Cardiac toxicity is a leading cause of preclinical safety failures in drug development. The cardiac markers NT-proANP and NT-proBNP have proven to be useful in preclinical toxicology testing.

Cardiac Safety Biomarker Assays in Preclinical Toxicology Testing

 

The BIOMEDICA NT-proANP and NT-proBNP ELISA kits are robust assays to quantify these cardiac hormones in rat samples. Due to the high inter-species homology the NT-proANP kit has also been applied in canine and feline samples.

BIOMEDICA´s NT-proANP ELISA kit (BI-20892) is widely published and has been independently validated for cardiovascular safety studies in rats (1, 2).

Rat NT-proBNP 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

 

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)

 

PRODUCT INFORMATION

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

 

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

 

CARDIAC SAFETY BIOMARKER ASSAYS in PRECLINICAL TOXICOLOGY TESTING 

References/Citations/Related Literature

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 Jan-Feb;77:58-65. doi: 10.1016/j.vascn.2015.10.002. Epub 2015 Oct 26. PMID: 26516096.
2. Natriuretic Peptides as Cardiovascular Safety Biomarkers in Rats: Comparison With Blood Pressure, Heart Rate, and Heart Weight. Engle SK, Watson DE.Toxicol Sci. 2016 Feb;149(2):458-72. doi: 10.1093/toxsci/kfv240. Epub 2015 Nov 25. PMID: 26609138.

 

Abstract

When using an ELISA assay, it is important to ensure that the results are specific, accurate, sensitive, and reproducible. The ELISA assay reliability  varies among kit suppliers, so choosing the “right” ELISA requires careful consideration. Access to the assay´s validation data before making a choice may be helpful.

ELISA Assay Reliability

At BIOMEDICA we take pride in developing and manufacturing our ELISA kits following a stringent manufacturing and quality control process, that enables us to maintain consistent and reproducible outcomes with every lot we produce.

At Biomedica we develop and manufacture our ELISA assays with care

 

All BIOMEDICA kits are fully validated and come with sample data for healthy human subjects, ready-to-use standards and controls.  We supply our kits with color-coded vials, and some of the reagents are  “colorful” as well, to make the kits easy to use and to avoid possible pipetting errors, To increase transparency, we publish the validation data of every ELISA assay kit on our website. 

BIOMEDICA´s  QUALITY GUIDELINES

AT BIOMEDICA, our goal is to supply reliable and well-validated ELISA kits for your research.

Here is how we ensure product excellence:

1. Optimization: we diligently optimize all Biomedica assay to guarantee reliability, sensitivity, and precision.
2. Validation: our kits undergo an extensive validation process in accordance with international quality guidelines (FDA, EMA, and ICH), ensuring the kits accuracy and efficiency.
3. Expert Team: our dedicated team consists of highly qualified scientists, many with doctorate-level education and industry training. Our team has extensive research experience and contributes to the development, production, and customer service aspects of our products.
4. Quality Management: Biomedica adheres to the ISO 9001: 2015 certified quality management system in our manufacturing process, ensuring consistent and high-quality products.

 

With these measures in place, we are committed to deliver ELISA assays that meet the highest standards of performance and reliability.

Learn more: https://www.bmgrp.com/quality 

Related literature

Interference in ELISA. Matson RS. 2023. Methods Mol Biol. 2612:91-99. doi: 10.1007/978-1-0716-2903-1_7. PMID: 36795361.

Abstract

Practical Guide to Immunoassay Method Validation. Andreasson U et al. 2015. Front Neurol. 19;6:179. doi: 10.3389/fneur.2015.00179. PMID: 26347708; PMCID: PMC4541289.

 

EZ4U –  Easy and Reliable Assay for Cell Viability Analysis

Cell viability quantifies the percentage of living and healthy cells within a given cell population. Cell viability analysis assays are employed to assess cell survival for instance after treatment with substances during drug screening.

Testing of Cell Viability with Biomedica´s EZ4U Assay

The EZ4U assay is a metabolic assay (like the MTT assay) that quantifies cell health by measuring the reduction of the colorimetric substrate through the activity of mitochondrial enzymes.

Cell viability assays are widely used in cell biology research. The EZ4U test kit is a reliable and straightforward non-radioactive assay that can be completed within two to five hours, depending on the cell type studied. The EZ4U assay employs non-toxic tetrazolium salts, which are converted into colored formazan. As the reduction process relies on functional mitochondria that become inactive shortly after cell death, this method effectively distinguishes between living and dead cells. Furthermore, as the assay procedure is identical to thymidine incorporation procedure, no changes in test protocols are necessary. An additional benefit is the ability to continue cultivation after determining cell numbers.

The EZ4U cell proliferation and cytotoxicity assay is highly suited for testing cell viability in a number of different cell types. For examples please follow the citations by clicking this link.

EZ4U Cell Proliferation and  Cytotoxicity Assay –straightforward with a single incubation step using living cells

The EZ4U assay (Biomedica, Austria) was used in a recent study investigating the effects of RNA methylation, namely m⁶A methylation, on the renal cell carcinoma (ACHN and 769P) cell lines. Renal cell carcinoma (RCC) accounts for about 2% of cancer-related deaths. Patients with metastatic RCC have a poor prognosis with an approximate 5% survival rate. Despite therapeutical advancements, including checkpoint immunotherapy,  the improvement in patient survival rates has been rather moderate.

Learn more: Depletion of the m⁶A demethylases FTO and ALKBH5 impairs growth and metastatic capacity through EMT phenotype change in clear cell renal cell carcinoma. Hu W, Klümper N, Schmidt D, Ritter M, Ellinger J, Hauser S. Am J Transl Res. 2023 Mar 15;15(3):1744-1755. PMID: 37056835; PMCID: PMC10086911.

Abstract

Background: N⁶-methyladenosine (m⁶A) is one of the most common RNA modifications in eukaryotes and has effects on RNA structure and stability. Recent studies have shown that m⁶A methylation is involved in human carcinogenesis. In the present study, we investigated the effects of m⁶A demethylases FTO and ALKBH5 on renal cell carcinoma (RCC) cell lines. Methods: The epithelial-mesenchymal in vitro knockdowns of FTO and ALKBH5 induced by antisense oligonucleotides (LNA-GapmeR system) were established in RCC cell lines. Their effects on migration and proliferation were investigated subsequently. The influence of FTO and ALKBH5 knockdown on key epithelial-mesenchymal transition (EMT) genes was analyzed. Results: Inactivation of FTO and ALKBH5 resulted in decreased proliferation and motility in all cell lines examined (ACHN, Caki-1, 769-P). Vimentin (VIM) was downregulated after the knockdown of FTO and ALKBH5, indicating an EMT switch. Conclusions: Knockdown of the m⁶A erasers FTO and ALKBH5 inhibits the malignant potential in the cell cultures studied by means of an EMT switch.

EZ4U  Cell Proliferation & 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 240 publications

 

Find out more: BROCHURE – EZ4U cell proliferation and cytotoxicity assay

 

 

 

 

At Biomedica we develop and manufacture quality ELISAs to quantify cytokine release

Our ELISAs to quantify cytokine release are validated according to international quality guidelines to ensure their consistency, specificity, precision, and robustness. The validation data files can be found on the respective product pages www.bmgrp.com. 

Our kits are designed to specifically quantify cytokine release in various biological matrices (e.g. serum, plasma, cell culture).

WHO reference reagents for a harmonized standardization

All our cytokine kits are standardized using WHO reference reagents/standards to allow cross-comparison of results when using different reagent sets during a study (1).

BIOMEDICAs CYTOKINE ELISA KITS

 

IL-6 ELISA (cat. no. BI-IL6)

The Biomedica IL-6 (interleukin-6) ELISA kit is a sandwich ELISA that incorporates two epitope-mapped antibodies that specifically bind to human IL-6 in the respective samples.

Capture antibody (pre-coated on a 96-well microtiter plate): recombinant IL-6 antibody (specific for human IL-6).

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

• • Method: sandwich ELISA
  • Sample type: serum, plasma, cell-culture, urine
  • Sample volume: 100µl / well
  • Assay time: 4.5 hrs
  • Standard range: 0 / 3.1 – 200 pg/ml
  • Sensitivity: LOD: 0.28 pg/ml; LLOQ: 0.78 pg/ml (measurable concentrations in serum AND plasma samples!)
  • Precision: in-between-run (n=9 ): ≤ 6 % CV, within-run (n=3): ≤ 7 % CV
  • Specificity:  the ELISA recognizes recombinant and endogenous (natural) human IL-6
  • Standard matrix: serum based matrix containing recombinant IL-6 including 8x vials of pre-diluted standards, lyophilized
  • Controls: 2 controls (high and low) included

 

VEGF ELISA (cat. no. BI-VEGF)

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

Capture antibody (pre-coated on a 96-well microtiter plate): recombinant VEGF antibody (specific for human VEGF) recognizes a structural epitope in the conserved receptor binding- site of the VEGF peptide. The antibody binds to all VEGF isoforms.

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

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

 

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

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

Capture antibody (pre-coated on a 96-well microtiter plate): recombinant ANG2 antibody (specific for human ANG2).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: polyclonal ANG2 antibody (streptavidin-HRPO labeled), specific for human ANG2, recognizing several linear epitopes that are distributed over the entire Angiopoietin-2 molecule.  

• Method: sandwich ELISA
• Sample type: serum, plasma, cell-culture, urine
• Sample volume: 20µl / well
• Assay time: 3.5 hrs
• Standard range: 0 / 12.5 – 400 pg/ml
• Sensitivity: 3.7 pmol/l (= 203 pg/ml)
• Precision: In-between-run (n=9): ≤ 6 % CV, Within-run (n=3): ≤ 8 % CV
• Specificity: the ELISA recognizes recombinant and endogenous (natural) human ANG2.  The assay most probably detects all three ANG2 isoforms, as determined by epitope mapping and analysis of the ANG2 sequence. ANG1 is not detected in this ELISA assay.
• Standard matrix: Serum based matrix containing recombinant ANG2,  including 8x vials of pre-diluted standards, lyophilized
• Controls: 2 controls (high and low) included

 

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

 

Related publications

1.       International reference reagents 

2.       Targeting IL-6 trans-signalling: past, present and future prospects. Rose-John S, Jenkins BJ, Garbers C, Moll JM, Scheller J. Nat Rev Immunol. 2023 Apr 17:1–16. doi: 10.1038/s41577-023-00856-y. Epub ahead of print. PMID: 37069261; PMCID: PMC10108826.

Abstract

Interleukin-6 (IL-6) is a key immunomodulatory cytokine that affects the pathogenesis of diverse diseases, including autoimmune diseases, chronic inflammatory conditions and cancer. Classical IL-6 signalling involves the binding of IL-6 to the membrane-bound IL-6 receptor α-subunit (hereafter termed ‘mIL-6R’) and glycoprotein 130 (gp130) signal-transducing subunit. By contrast, in IL-6 trans-signalling, complexes of IL-6 and the soluble form of IL-6 receptor (sIL-6R) signal via membrane-bound gp130. A third mode of IL-6 signalling – known as cluster signalling – involves preformed complexes of membrane-bound IL-6-mIL-6R on one cell activating gp130 subunits on target cells. Antibodies and small molecules have been developed that block all three forms of IL-6 signalling, but in the past decade, IL-6 trans-signalling has emerged as the predominant pathway by which IL-6 promotes disease pathogenesis. The first selective inhibitor of IL-6 trans-signalling, sgp130, has shown therapeutic potential in various preclinical models of disease and olamkicept, a sgp130Fc variant, had promising results in phase II clinical studies for inflammatory bowel disease. Technological developments have already led to next-generation sgp130 variants with increased affinity and selectivity towards IL-6 trans-signalling, along with indirect strategies to block IL-6 trans-signalling. Here, we summarize our current understanding of the biological outcomes of IL-6-mediated signalling and the potential for targeting this pathway in the clinic.

 

Fibroblast growth factor 23 (FGF23) is a hormone that plays a crucial role in regulating serum phosphate and vitamin D levels within the body. FGF23 is primarily produced in the bone by osteocytes and osteoblasts in response to factors such as oral phosphate intake or elevated serum Vitamin D concentrations. The regulation of normal serum phosphorus levels is primarily achieved through a highly controlled process occurring in the kidney where phosphate reabsorption takes place. Patients suffering from chronic kidney disease (CKD) have high plasma FGF23 levels and FGF23 serves as a sensitive biomarker for detecting abnormal renal phosphate handling. Notably, FGF23 levels increase during the early stages of kidney dysfunction, providing valuable insights into the underlying renal impairments (1, 2).

Effects of FGF23 on the heart

High FGF23 concentrations have been found to be associated with multiple cardiac diseases, including left ventricular hypertrophy, heart attacks, heart failure, and cardiovascular related deaths (3-5).

Present findings exploring the relationship of FGF23 and cardiac events are presented in a recent review. The authors further discuss the potential mechanisms by which FGF23 directly or indirectly triggers left ventricular hypertrophy (6). 

Learn more: Direct and indirect effects of fibroblast growth factor 23 on the heart. Nakano T et al., 2023.

Download our leaflet “FGF23 – an overview” here

How can FGF23 be measured?

Circulating FGF23 concentrations can be measured through blood tests by immunoassays. A widely used method for measuring FGF23 is with an ELISA assay (enzyme-linked immunosorbent assays). FGF23 ELISA assays utilize specific antibodies that recognize and bind FGF23 present in the sample.

Currently, there are two different assays commercially available for measuring circulating FGF23 in blood samples (7).

Intact FGF23 Assays

intact FGF23 (iFGF23) represents the full length, biologically active form of the hormone. It consists of the complete FGF23 protein structure without being enzymatically cleaved. The two antibodies in these assays target the N-terminal part and the C-terminal domain of the FGF23 molecule, respectively.

FGF23 C-terminal Assays

The c-terminal fragments of FGF23 (cFGF23) are the result of the enzymatic cleavage of the intact FGF23 molecule. The antibodies utilized in the cFGF23 assays bind to specific epitopes that are located in the c-terminal domain of the FGF23 molecule.

Noteworthy, all commercially available FGF23 c-terminal assays detect both c-terminal FGF23 fragments as well as the intact FGF23 molecule (7).

BIOMEDICA has developed two distinct ELISA assays to reliably quantify FGF23 concentrations in human serum and plasma.

·       FGF23 intact ELISA (cat. no. BI-20700)

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

• Validated FGF23 ELISA kits according to international quality guidelines
• Numerous references in top-ranking journals

 

All Assays are Developed & Manufactured by Biomedica

Literature

1. The regulation of FGF23 under physiological and pathophysiological conditions. Rausch S, Föller M. Pflugers Arch. 2022 Mar;474(3):281-292. doi: 10.1007/s00424-022-02668-w. Epub 2022 Jan 27. PMID: 35084563; PMCID: PMC8837506.
2. Regulation and Effects of FGF23 in Chronic Kidney Disease. Musgrove J, Wolf M. Annu Rev Physiol. 2020 Feb 10;82:365-390. doi: 10.1146/annurev-physiol-021119-034650. Epub 2019 Nov 19. PMID: 31743079.
3. FGF23 and Cardiovascular Risk. Prié D. Ann Endocrinol (Paris). 2021 Jun;82(3-4):141-143. doi: 10.1016/j.ando.2020.03.007. Epub 2020 Mar 10. PMID: 32950228.
4. FGF23 predicts outcomes in heart failure but questions remain unanswered. Duran A, daSilva-deAbreu A, Joury A, Ventura HO. Int J Cardiol. 2021 Sep 1;338:145-146. doi: 10.1016/j.ijcard.2021.06.036. Epub 2021 Jun 19. PMID: 34157358.
5. Paracrine Effects of FGF23 on the Heart. Front Endocrinol (Lausanne). Leifheit-Nestler M, Haffner D 2018 May 28;9:278. doi: 10.3389/fendo.2018.00278. PMID: 29892269; PMCID: PMC5985311.
6. Direct and indirect effects of fibroblast growth factor 23 on the heart. Nakano T, Kishimoto H, Tokumoto M Front Endocrinol (Lausanne). 2023 Feb 24;14:1059179. doi: 10.3389/fendo.2023.1059179. PMID: 36909314; PMCID: PMC9999118.
7. The Measurement and Interpretation of Fibroblast Growth Factor 23 (FGF23) Concentrations. Calcif Tissue Int. Heijboer AC, Cavalier E. 2023 Feb;112(2):258-270. doi: 10.1007/s00223-022-00987-9. Epub 2022 Jun 4. PMID: 35665817; PMCID: PMC9859838.

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• Reliable Assay Kit
• Easy, Reliable NT-ProANP ELISA Kit
• Easy-to-Use System that Delivers the Desired Results
• Measuring Cell Proliferation with EZ4U

 

 

YOUR SATISFACTION IS OUR PRIORITY

Our goal is to meet your needs at every stage, starting from assay design and development, through manufacturing, validation and customer support. We have implemented a range of measures to achieve this:

• Ready-to-Use Assays
• Biomarker Detection in Real Samples
• Validation According to International Standards
• Qualified, Experienced Customer Service
• Flexibility
• Service Measurements

 

learn more “About Biomedica” 

QUALITY is not just a word

Rest assured when using our products – we prioritize quality throughout the entire process from product development to manufacturing and quality control. Our goal is to ensure that the performance of our products meet the top quality standards.

Right after product development our kits go through a rigorous validation process in compliance with the international quality guidelines (FDA, EMEA, ICH).  Each product is accompanied by a comprehensive validation report that can downloaded from our website www.bmgrp.com .

WE ARE HERE FOR YOU

The vast majority of Biomedica products is delivered and used without any issues. However, if you do encounter a problem or have a question, we are here to help. Our dedicated and qualified team will do their best to answer your questions and will be there for you. Due to the close collaboration between our team members in development and production, we are in a unique position to support our customers with all of the detailed information they may need.

Being scientists at heart, we are always happy to share our knowledge, give advice, and help our customers selecting biomarkers.

 

Role of C-Type Natriuretic Peptide (CNP) in bone growth and development

CNP is a member of the natriuretic peptide family. It acts as a paracrine hormone which is present in several tissues such as the cardiovascular system, brain and the endothelium. CNP controls various processes including blood pressure, water and mineral balance, and numerous metabolic functions (1). In 1995, CNP was first described as a potent regulator of bone remodeling, underlying its important role in humans (2).

C-Type Natriuretic Peptide Induces Bone Growth

Research has indicated that CNP is predominantly synthesized within the joint cartilage by chondrocytes, which are specialized cells primarily responsible for generating collagen and the extracellular matrix. As a result, the primary and well-established role of CNP is in the growth plate skeletal cartilage, where it plays a crucial role in promoting endochondral growth of the developing skeleton (3, 4).

What is the mechanism of bone growth?

The growth of bones is a complex process involving the formation and remodeling of bone tissue throughout the body. It is regulated by a diverse range of hormones, growth factors, and signaling pathways that interact together to maintain skeletal health and function.  

Bone growth can be broadly classified into two types:

Longitudinal growth primarily takes place in the long bones of the body, such as femur and tibia. It is a process by which bones grow in length, predominantly occurring during childhood and adolescence. Longitudinal growth is facilitated by growth plates, which are cartilaginous areas located at the ends of long bones. As the bones elongate, the cartilage in the growth plates gradually undergoes ossification, transforming into bone tissue.

 Appositional growth, on the other hand, is the process by which bones grow in thickness or diameter. This type of growth occurs throughout life and is regulated by a delicate balance between osteoblasts, cells responsible for bone formation and osteoblasts, cells involved in bone resorption and bone-resorbing cells named osteoclasts.

Numerous factors can influence bone growth, including genetics, nutrition, exercise, and hormonal equilibrium. Hormones such as growth hormone and sex hormones play crucial roles in regulating bone growth and development. One hormone that is of particular significance in this regard is C-type natriuretic peptide (CNP), which is essential for the regulation of bone growth.

Quantification of CNP and NT-proCNP in human blood samples

The measurement of biologically active CNP in human blood samples presents a challenge due to its dynamic role in bone growth, its rapid clearance and low circulating concentrations. However, the inactive, but more stable precursor molecule NT-proCNP, has proven to be a reliable alternative. The amino-terminal propeptide NT-proCNP, is synthesized in the growth plate, has a longer half-life and circulates in equimolar concentrations as CNP. Therefore, NT-proCNP is an excellent candidate for investigating the role of C-Type natriuretic Peptide in bone (1, 3).

A study comparing the measurement of CNP and NT-proCNP in human blood samples revealed that an NT-proCNP assay provides more consistent and reliable results, making it the preferred choice for clinical applications (5).

NT-proCNP can reliably be measured in serum and plasma samples with a conventional ELISA assay 

 BIOMEDICA – NT-proCNP ELISA (cat. no BI-20812) 

• Full validation  – following international quality guidelines
• For human and non-human samples (serum, plasma, cell culture, urine)
• Standard 96-well ELISA assay format- color coded and ready to use reagents
• Developed & manufactured by Biomedica  

 

NT-proCNP serves as a biomarker of linear growth  

Research studies in children has provided evidence showing a strong correlation between the levels of NT-proCNP in blood and the rate of linear growth throughout the entire growth process. In infancy, both males and females exhibit high levels of CNP and NT-proCNP, which subsequently decline in early childhood. However, during puberty, NT-proCNP levels rise again, followed by a decrease during adulthood. The peak levels of NT-proCNP align with the age at which peak height velocity occurs. As a result, the measurement of CNP syntheses through NT-proCNP in blood is highly indicative of linear growth in healthy children at all stages of development, making it an exceptional biomarker for monitoring linear growth (3, 6).

The role of NT-proCNP in bone diseases and genetic disorders

While the involvement of CNP in bone growth during development is well-established, its precise role in the adult skeleton remains uncertain. It is plausible that bone remodeling may contribute to increased plasma levels of CNP in children, given that CNP is expressed in osteoblasts and osteoclasts, which are the cells responsible for bone formation and bone degradation. Moreover, alternations in plasma CNP products have been linked to changes in bone turnover markers in adults (7),

CNP has emerged as a significant player in various types of skeletal dysplasia, a group of over 400 genetic disorders characterized by abnormal bone, joint, and cartilage development. Mutations in genes associated with CNP signaling have been identified in these conditions, including achondroplasia and hypochondroplasia, which are the most prevalent forms of dwarfism (8).

CNP has shown to be a promising therapeutic target for treating growth and skeletal disorders. Only recently, a variant of CNP was approved for treatment of achondroplasia (9). Ongoing clinical trials are currently investigating additional inhibitors of the CNP signaling pathway (10).

Related ELISA assay products

-NT-proANP (cat. no. BI-20492) – assay also suitable for preclinical applications (rat, mouse and other species) – 10µl sample volume

-NT-proBNP (cat. no. SK-1204) – CE-marked, for IVD use in EU

-NT-proBNP rat (cat. no. BI-1204R) – sample values provided

 

Literature

1. Circulating products of C-type natriuretic peptide and links with organ function in health and disease. Prickett TC, A Espiner E. Peptides. 2020. 132:170363.
2. C-type natriuretic peptide increases bone resorption in 1,25-dihydroxyvitamin D3-stimulated mouse bone marrow cultures. Holliday LS, Dean AD, Greenwald JE, Glucks SL. J Biol Chem. 1995. 11;270(32):18983-9.
3. Plasma C-Type Natriuretic Peptide: Emerging Applications in Disorders of Skeletal Growth. Espiner E, Prickett T, Olney R. Horm Res Paediatr. 2018;90(6):345-357.
4. Hormones and osteoporosis update. Effects of natriuretic peptides on endochondral bone growth. Clin Calcium. 2009. 19(7):1003-8. Yasoda A, Nakao K. Japanese. PMID: 19567998.
5. Comparative measurement of CNP and NT-proCNP in human blood samples: a methodological evaluation. Kuehnl A, Pelisek J, Bruckmeier M, Safi W, Eckstein HH. J Negat Results Biomed. 2013. 1;12:7.
6. Amino-terminal propeptide of C-type natriuretic peptide (NTproCNP) predicts height velocity in healthy children. Olney RC, Permuy JW, Prickett TC, Han JC, Espiner EA. Clin Endocrinol (Oxf). 2012. 77(3):416-22.
7. C-type natriuretic peptide forms in adult hyperthyroidism: correlation with thyroid hormones and markers of bone turnover. Schouten BJ, Prickett TC, Hunt PJ, Richards AM, Geffner ME, Olney RC, Espiner EA. Clin Endocrinol (Oxf). 2012. 76(6):790-6.
8. New developments in the management of achondroplasia. Högler W, Ward LM. Wien Med Wochenschr. 2020. 170(5-6):104-111.
9. Expanding horizons of achondroplasia treatment: current options and future developments. Fafilek B, Bosakova M, Krejci P. Osteoarthritis Cartilage. 2022. 30(4):535-544.
10. Emerging drug targets for achondroplasia. Savarirayan R. Expert Opin Ther Targets. 2022. 26(5):389-391.

 

 

 

 

We are happy to introduce our new brochure featuring our Quality Cytokine Assays “BIOMEDICA´s CYTOKINE ELISA kits”  

Product Highlights

• Full validation package
• Ready-to-use prediluted standards
• Color coded reagents & controls included
• Kit calibration with WHO international standard

QUALITY CYTOKINE ELISA KITS FOR AFFORDABLE & EFFICIENT RESEARCH

human Cytokine ELISA Kits  – developed & manufactured by Biomedica

• IL-6 (cat. no. BI-IL6) –  high sensitivity – detectable levels
• VEGF (cat. no. BI-VEGF) –  low sample volume – 10 µl – detecting all known VEGF isoforms
• ANGIOPOIETIN-2 (cat. no. BI-ANG2)    / optimized assay range 

 

Also available mouse/rat Angiopoietin-2 ELISA kit (cat. no. BI-ANG2MR) –  only 5µl sample volume.

 

 

ABOUT CYTOKINES

Cytokines are critical cell signaling molecules that play important roles in the immune response and many other physiological processes.  These proteins are produced by a wide variety of cell types that act on specific target cell receptors to trigger a range of biological responses.

Cytokines are involved in many aspects of the immune response, including inflammation, cell growth and differentiation, and cell death (apoptosis). Cytokines also regulate tissue repair and wound healing. Dysregulation of cytokine production and signaling may contribute to various diseases including autoimmune disorders, allergies, and cancer.

Some common cytokines include interleukins, tumor necrosis factor (TNF), and interferons.

Interleukin-6 (IL-6) is a pleiotropic cytokine that is largely involved in immunomodulatory processes. One of its roles is to respond to infections. As a pro-inflammatory cytokine, IL-6 activates the immune response, recruiting immune cells triggering B and T cell response. IL-6 dysregulation is associated with pathologies involving chronic inflammation, autoimmune diseases, and atherosclerosis. However, IL-6 also has beneficial anti-inflammatory and positive metabolic effects that are released during exercise coupled to a physical adaptation to intense training. In addition to its immune functions, IL-6 is involved in many other physiological processes, including bone remodeling and the nervous system.

Angiopoietin-2 (ANG2) is a protein that plays a critical role in regulating the development and remodeling of blood vessels, a process known as angiogenesis. ANG-2 is primarily produced by endothelial cells, which are cells that line the interior of blood vessels. Under normal physiological conditions the expression of ANG2 is low, which allows the blood vessels to undergo a rapid growth and remodeling. However, in pathological conditions, such as inflammation and cancer, ANG2 expression can be upregulated, leading to excessive angiogenesis and blood vessel destabilization.

Vascular endothelial growth factor (VEGF) is a protein mainly produced by endothelial cells. VEGF stimulates the growth and proliferation of endothelial cells, as well as the migration and differentiation of these cells into new blood vessels. VEGF plays a role in the development of cancer and certain ocular diseases, where it promotes the growth of new blood vessels that feed tumors or damage the retina.

There are several different isoforms of VEGF, each with different properties and functions. For example, VEGF-A is the most well-known cytokine of the VEGF family, and is primarily responsible for promoting angiogenesis. Other isoforms, such as VEGF-B and VEGF-C, have different roles in the body, such as regulating the growth of blood vessels in the heart and lymphatic system, respectively.

Related publications

Harnessing cytokines and chemokines for cancer therapy. Propper DJ, Balkwill FR. Nat Rev Clin Oncol. 2022 Apr;19(4):237-253. doi: 10.1038/s41571-021-00588-9. Epub 2022 Jan 7. PMID: 34997230.

Inflammatory Cytokines and Chemokines as Therapeutic Targets in Heart Failure. Hanna A, Frangogiannis NG. Cardiovasc Drugs Ther. 2020 Dec;34(6):849-863. doi: 10.1007/s10557-020-07071-0. Epub 2020 Sep 9. PMID: 32902739; PMCID: PMC7479403.

Around 7 out of 100,000 people are affected by primary malignant brain tumors. Glioblastoma multiforme (GBM) is the most common type of brain tumor. It grows rapidly and has a low prognosis of approximately 15 months. The  mean age after diagnosis is 63 years (1, 2). Despite significant progress in treatment, there remains a critical need for novel treatment options for GBM. 

Natural Malaria Substance – novel treatment option for brain tumors

Artemisinin, deriving from the plant Artemisia annua is a potent natural substance globally used to treat Malaria. It was discovered in the 1970s from Traditional Chinese Medicines and has since then saved millions of lives (3). Due to its cytotoxic properties it has not only been acknowledged for treating Malaria but also for its potential anticancer properties. With the discovery of Artemisinin and its derivates, researchers have been investigating the mechanisms on how these compounds work e.g. in inhibiting tumor cell proliferation, promoting apoptosis, disrupting caner invasion and metastasis, and preventing angiogenesis (4).

The natural malaria substance Artemisinin  –  a novel treatment option for brain tumors. 

In a recent study using genome-wide screening, researchers have identified mitochondrial function, particularly the biosynthesis of porphyrin, as a crucial pathway responsible for Artemisinin´s cytotoxicity. The potential of this novel treatment option for human brain tumors is currently under investigation (5, 6).

EZ4U / MTT Cell viability assay (cat. no. BI-5000) – BIOMEDICA

 

The Biomedica EZ4U cell viability and cytotoxicity assay was employed in mammalian cell lines in the described study:

A whole-genome scan for Artemisinin cytotoxicity reveals a novel therapy for human brain tumors.

The non-toxic and non-radioactive assay EZ4U / MTT Cell viability assay is based on the conversion of the yellow tetrazolium salt to coloured soluble formazan, which is produced by mitochondrial enzymes. The quantity of formazan produced is in direct proportion to the number of viable, living cells present.

Assay Characteristics

• Method: Reduction of tetrazolium salt to coloured formazan
• Sample type cell culture medium
• Sample size 200 μl / test, 10×96 tests
• Detection limit depending on cell lines Incubation time 2 – 5 h

 

Click here to download our EZ4U “easy for you” – brochure

Literature

1. Brain Tumors. Am J Med. McFaline-Figueroa JR, Lee EQ. 2018 Aug;131(8):874-882. doi: 10.1016/j.amjmed.2017.12.039. Epub 2018 Jan 31. PMID: 29371158.
2. Rising Incidence of Glioblastoma Multiforme in a Well-Defined Population. Grech N, Dalli T, Mizzi S, Meilak L, Calleja N, Zrinzo A Cureus. 2020 May 19;12(5):e8195. doi: 10.7759/cureus.8195. PMID: 32572354; PMCID: PMC7302718.
3. The discovery of artemisinin and the Nobel Prize in Physiology or Medicine. Su XZ, Miller LH. Sci China Life Sci. 2015 Nov;58(11):1175-9. doi: 10.1007/s11427-015-4948-7. PMID: 26481135; PMCID: PMC4966551.
4. Antitumor Research on Artemisinin and Its Bioactive Derivatives. Zhang Y, Xu G, Zhang S, Wang D, Saravana Prabha P, Zuo Z. Nat Prod Bioprospect. 2018 Aug;8(4):303-319. doi: 10.1007/s13659-018-0162-1. Epub 2018 Apr 9. PMID: 29633188; PMCID: PMC6102173.
5. A whole-genome scan for Artemisinin cytotoxicity reveals a novel therapy for human brain tumors. Taubenschmid-Stowers J, Orthofer M, Laemmerer A, Krauditsch C, Rózsová M, Studer C, Lötsch D, Gojo J, Gabler L, Dyczynski M, Efferth T, Hagelkruys A, Widhalm G, Peyrl A, Spiegl-Kreinecker S, Hoepfner D, Bian S, Berger W, Knoblich JA, Elling U, Horn M, Penninger JM. EMBO Mol Med. 2023 Mar 8;15(3):e16959. doi: 10.15252/emmm.202216959. Epub 2023 Feb 6. PMID: 36740985.
6. Achilles heel of glioblastoma. JLP Health and its partners discover combinatorial treatment option for incurable brain tumors. Press Release, JLP Health GmbH, Vienna, Austria, 7 February 2023.

Bone metastases affect over 1.5 million cancer patients globally, making bones a favored metastatic site for solid tumors (1). The presence of skeletal metastases can significantly reduce the quality of life for individuals with advanced cancer, as weakened bones can cause pain, fractures, and increase the risk of mortality (2, 3).

Bones have numerous important functions which include structural support, mobility, hematopoiesis, and mineral storage (4). The health and performance of our bone tissue are overseen by a range of cell types, which include:

• Osteoblasts – cells responsible for creating new bone tissue
• Osteoclasts – cells responsible for breaking down bone tissue
• Osteocytes –  cells within the bone that monitor mechanical loading and regulate the process of bone remodeling 

 

Bone biology- cancer and bone

Cancer and Bone – bone a preferred target site for cancer metastasis

Bone is a favored destination for metastasis in specific types of cancer, whereby cancer cells detach from the primary tumor and disseminate throughout the body. Certain cancers, such as breast, prostate, kidney, lung, ovarian, and thyroid, are especially prone to spread to bone (5).

Cancer and  Bone – the RANK/RANKL/OPG system

The RANK/RANKL/OPG system (receptor activator of the nuclear factor-κB ligand/ Osteoprotegerin) was identified more than 20 years ago and remains a widely researched topic until this day. The interaction between RANK and RANKL is essential for bone metabolism and osteoclast development. OPG acts as a decoy receptor for RANKL. By binding completely to RANKL, OPG obstructs the RANKL-RANK interaction, thereby blocking bone resorption. In addition to its role in regulating bone remodeling, the RANK/RANKL/OPG system is directly implicated in tumor cell development, particularly in the progression of breast and prostate cancer as well as leukemia (6-8).

Inhibiting RANK/RANKL signaling in human cancer

A fully humanized monoclonal antibody has been developed to counteract the effects of RANKL, which has received approval for treating bone loss conditions. The antibody functions by disrupting RANK signaling, preventing osteoclast activation and inhibiting bone resorption (6-8). More recently, inhibiting RANKL has been recognized as a significant checkpoint with the potential to influence anti-tumor response. Consequently, RANKL blockade has a direct impact on bone metastasis. Ongoing clinical and experimental trials are evaluating this emerging therapeutic approach (9).

Studies analyzing serum levels of OPG and RANKL

Serum concentrations of both OPG and soluble RANKL can be accessed via an enzyme-linked immunosorbent assay (ELISA). The predictive value of RANKL/OPG serum levels and disseminated tumor cells in breast cancer patients without metastasis was investigated in 509 patients with primary, nonmetastatic breast cancer. The results demonstrated that RANKL serum levels were significantly elevated in patients who developed bone metastases (10). A different study highlighted the role of OPG as a marker of breast cancer risk in women with a BRCA1 mutation. The authors suggested that OPG levels could be associated with disease risk, potentially serving as a marker for breast cancer risk and improving existing risk prediction models by identifying women at high risk of developing the disease (11).

How are circulating serum levels of Osteoprotegerin and soluble RANKL measured?

Both markers can easily be measured with a conventional ELISA assay.

• Osteoprotegerin (OPG) ELISA (cat. no. BI-20403) – day test, 20µl sample volume/well
• Soluble free RANKL ELISA (cat. no. BI-20462) – highly sensitive and reliable

 

– Widely cited in + 450 publications (citations for OPG, and RANKL)

– Extensively validated ELISA assays following international quality guidelines

High Quality ELISA kits – Developed & Manufactured by Biomedica

Related ELISA kits:  DKK-1, Sclerostin, IL-6, VEGF, Angiopoietin-2

References

1. Biology of Bone Tissue: Structure, Function, and Factors That Influence Bone Cells. Florencio-Silva R, Sasso GR, Sasso-Cerri E, Simões MJ, Cerri PS. Biomed Res Int. 2015;2015:421746. doi: 10.1155/2015/421746. Epub 2015 Jul 13. PMID: 26247020; PMCID: PMC4515490.
2. Understanding the Bone in Cancer Metastasis. Fornetti J, Welm AL, Stewart SA. J Bone Miner Res. 2018 Dec;33(12):2099-2113. doi: 10.1002/jbmr.3618. Epub 2018 Nov 26. PMID: 30476357.
3. Bone as a Preferential Site for Metastasis. JBMR Plus. Sowder ME, Johnson RW. 2019 Jan 15;3(3):e10126. doi: 10.1002/jbm4.10126. PMID: 30918918; PMCID: PMC6419612.
4. Cancer to bone: a fatal attraction. Weilbaecher KN, Guise TA, McCauley LK. Nat Rev Cancer. 2011 Jun;11(6):411-25. doi: 10.1038/nrc3055. Epub 2011 May 19. PMID: 21593787; PMCID: PMC3666847.
5. Bone metastasis: mechanisms, therapies, and biomarkers. Clézardin P, Coleman R, Puppo M, Ottewell P, Bonnelye E, Paycha F, Confavreux CB, Holen I. Physiol Rev. 2021 Jul 1;101(3):797-855. doi: 10.1152/ physrev.00012.2019. Epub 2020 Dec 24. PMID: 33356915.
6. The Roadmap of RANKL/RANK Pathway in Cancer. Casimiro S, Vilhais G, Gomes I, Costa L. Cells. 2021 Aug 4;10(8):1978. doi: 10.3390/cells10081978. PMID: 34440747; PMCID: PMC8393235.
7. RANKL biology: bone metabolism, the immune system, and beyond. Ono T, Hayashi M, Sasaki F, Nakashima T. Inflamm Regen. 2020 Feb 7;40:2. doi: 10.1186/s41232-019-0111-3. PMID: 32047573; PMCID: PMC7006158.
8. RANKL and RANK in Cancer Therapy. Physiology (Bethesda). Onji M, Penninger JM. 2023 May 1;38(3):0. doi: 10.1152/physiol.00020.2022. Epub 2022 Dec 6. PMID: 36473204.
9. The Role of the RANKL/RANK Axis in the Prevention and Treatment of Breast Cancer with Immune Checkpoint Inhibitors and Anti-RANKL. Simatou A, Sarantis P, Koustas E, Papavassiliou AG, Karamouzis MV. Int J Mol Sci. 2020 Oct 14;21(20):7570. doi: 10.3390/ijms21207570. PMID: 33066388; PMCID: PMC7590202.
10. Prognostic Value of RANKL/OPG Serum Levels and Disseminated Tumor Cells in Nonmetastatic Breast Cancer. Rachner TD, Kasimir-Bauer S, Göbel A, Erdmann K, Hoffmann O, Browne A, Wimberger P, Rauner M, Hofbauer LC, Kimmig R, Bittner AK. Clin Cancer Res. 2019 Feb 15;25(4):1369-1378. doi: 10.1158/1078-0432.CCR-18-2482. Epub 2018 Nov 13. PMID: 30425091.
11. Delineating the role of osteoprotegerin as a marker of breast cancer risk among women with a BRCA1 mutation. Park SS, Uzelac A, Kotsopoulos J. Hered Cancer Clin Pract. 2022 Apr 13;20(1):14. doi: 10.1186/s13053-022-00223-3. PMID: 35418083; PMCID: PMC9008947.

 

We offer custom analytical testing services for the measurement of your samples. ANALYTICAL SERVICES – BIOMEDICA –  reliable and flexible:

METHODS:  ELISA  & Luminex Assays, microRNA Analysis, and Glycan Profiling.

Advantages of Using Biomedica Analytical Testing Services:

 

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

 

Check out our ELISA and Luminex Workflow chart and our microRNA Service Details 

Biomedica offers a broad range of high quality RNA services performed by experts according to GLP standards. These services include:

• 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.
• Biomedica and TAmiRNA 

Biomedica Analytical Services – reliable and flexible

We can measure any kind of selected biomarker on your “wish” list by ELISA (enzyme linked immunosorbent assay) for clinical and preclinical samples e.g.

• • Clinical samples (serum, plasma, urine, cell culture supernatants)
  • Pre-clinical samples (e.g. NT-proBNP or NT-proANP measurement in mouse, rat,  rabbit, or other species)  
  • Custom applications – contact us

 

Chronic Kidney Disease (CKD) affects more than 10% of the general population worldwide (1). It is characterized by kidney fibrosis, a progressive process that destroys the kidney (2). An effective treatment to stop the progression of CKD is still lacking. Scientists have now discovered a potential mean to treat CKD by inhibiting Angiopoietin-2 (ANG2), a vascular growth factor (3). Angiopoietin-2 is a potential new treatment target for kidney fibrosis in CKD.

Angiopoietin-2 a new treatment target for kidney fibrosis in CKD

Patients with chronic kidney disease (CKD) have increased plasma levels of Angiopoietin-2 (ANG2).  Studies have shown that ANG2 induces kidney injury and fibrosis in patients with CKD as well as in mouse models of kidney disease (3).  Preventing the progression of renal fibrosis by blocking Angiopoietin-2 could be a new approach in treating chronic kidney disease-associated pathologies.

Based on data of the recent study from Chang et al., on the application of an Angiopoietin-2 inhibitor to mouse models of progressive kidney disease, the scientists found a profound inhibitory effects of ANG2 inhibition on macrophage infiltration and fibrosis (3).  

Future clinical studies will be required to investigate the therapeutic potential on the protective effect of Angiopoietin-2 inhibition on progressive kidney disease. Read more: Angiopoietin-2 inhibition attenuates kidney fibrosis by hindering chemokine C-C motif ligand 2 expression and apoptosis of endothelial cells.

BIOMEDICA developed two assays for the accurate measurement of ANGIOPOIETIN-2 

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

• only 5µl sample volume /well
• no predilution of samples required
• kit control included
• sample values provided

 

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

• only 20µl sample volume /well
• the assay is optimized for clinical samples – sample values provided
• ready to use standards and controls  included

 

References

1. 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 Jun 30;20:100438. doi: 10.1016/j.lanepe.2022.100438. PMID: 36090671; PMCID: PMC9459126.

2. Fibrosis in Chronic Kidney Disease: Pathogenesis and Consequences. Panizo S, Martínez-Arias L, Alonso-Montes C, Cannata P, Martín-Carro B, Fernández-Martín JL, Naves-Díaz M, Carrillo-López N, Cannata-Andía JB. Int J Mol Sci. 2021 Jan 2;22(1):408. doi: 10.3390/ijms22010408. PMID: 33401711; PMCID: PMC7795409.

3. Angiopoietin-2 inhibition attenuates kidney fibrosis by hindering chemokine C-C motif ligand 2 expression and apoptosis of endothelial cells. Chang FC, Liu CH, Luo AJ, Tao-Min Huang T, Tsai MH, Chen YJ, Lai CF, Chiang CK, Lin TH, Chiang WC, Chen YM, Chu TS, Lin SL. Kidney Int. 2022 Oct;102(4):780-797. doi: 10.1016/j.kint.2022.06.026. Epub 2022 Aug 4. PMID: 35934136.

Abstract

Plasma levels of angiopoietin-2 are increased in patients with chronic kidney disease (CKD). Moreover, mouse models of progressive kidney disease also demonstrate increased angiopoietin-2 in both plasmas and kidneys. The role of dysregulated angiopoietins in the progression of kidney disease has not been thoroughly investigated. Here, we found in a cohort of 319 patients with CKD that plasma angiopoietin-2 and angiopoietin-2/angiopoietin-1 ratios were positively associated with the development of kidney failure. In mice with progressive kidney disease induced by either ureteral obstruction or ischemia-reperfusion injury, overexpression of human angiopoietin-1 in the kidney tubules not only reduced macrophage infiltration in the initial stage post-injury but also attenuated endothelial cell apoptosis, microvascular rarefaction, and fibrosis in the advanced disease stage. Notably, angiopoietin-1 attenuated chemokine C-C motif ligand 2 (CCL2) expression in the endothelial cells of the fibrosing kidneys, and these protective effects led to attenuation of functional impairment. Mechanistically, angiopoietin-1 reduced CCL2-activated macrophage migration and protected endothelial cells against cell apoptosis induced by angiopoietin-2 and Wnt ligands. Based on this, we applied L1-10, an angiopoietin-2 inhibitor, to the mouse models of progressive kidney disease and found inhibitory effects on macrophage infiltration, microvascular rarefaction, and fibrosis. Thus, we defined the detrimental impact of increased angiopoietin-2 on kidney survival of patients with CKD which appears highlighted by angiopoietin-2 induced endothelial CCL2-activated macrophage infiltration and endothelial cell apoptosis in their kidneys undergoing fibrosis.

Further reading

New therapeutic targets in chronic kidney disease progression and renal fibrosis. Rayego-Mateos S, Valdivielso JM. Expert Opin Ther Targets. 2020 Jul;24(7):655-670. doi: 10.1080/14728222.2020.1762173. Epub 2020 May 18. PMID: 32338087.

 

 

The International Day of Happiness has been initiated by the United Nations to spread awareness of the importance of happiness in the lives of people around the world (1). Happiness and health are closely tied together and affect one another.

Happiness and Health

Numerous studies have found that happiness is linked to a strong immune system and a better general health (2). Reducing risk factors  e.g., tobacco, alcohol, low-nutrition foods and beverages are only some strategies to reduce disease burden (3).

Studying biomarkers may help to understand the mechanism on how the disease originates and could improve diagnosis and disease prognosis (4).

BIOMEDICA specializes on the development of cutting-edge biomarker assays for clinical research.

Biomedica ELISA kits by research area

• Bone Metabolism
• Cardiovascular
• Cell Proliferation
• Immunology
• Infectious Disease
• Metabolic Disease
• Infection
• Nephrology and Transplant
• Neurology
• Oncology
• Oxidative Stress
• Cytokines

High quality ELISA kits- developed and manufactured by Biomedica

References

1. International Day of Happiness – the United Nations  
2. Happiness and Health Annu Rev Public Health. Steptoe A. 2019 Apr 1;40:339-359. doi: 10.1146/annurev-publhealth-040218-044150. Epub 2019 Jan 2. PMID: 30601719. Full text review

Abstract

Research into the relationship between happiness and health is developing rapidly, exploring the possibility that impaired happiness is not only a consequence of ill-health but also a potential contributor to disease risk. Happiness encompasses several constructs, including affective well-being (feelings of joy and pleasure), eudaimonic well-being (sense of meaning and purpose in life), and evaluative well-being (life satisfaction). Happiness is generally associated with reduced mortality in prospective observational studies, albeit with several discrepant results. Confounding and reverse causation are major concerns. Associations with morbidity and disease prognosis have also been identified for a limited range of health conditions. The mechanisms potentially linking happiness with health include lifestyle factors, such as physical activity and dietary choice, and biological processes, involving neuroendocrine, inflammatory, and metabolic pathways. Interventions have yet to demonstrate substantial, sustained improvements in subjective well-being or direct impact on physical health outcomes. Nevertheless, this field shows great potential, with the promise of establishing a favorable effect on population health.

3. Association of Healthy Lifestyle With Years Lived Without Major Chronic Diseases.Nyberg ST, Singh-Manoux A, Pentti J, .. Batty GD, Kivimäki M. JAMA Intern Med. 2020 May 1;180(5):760-768. doi: 10.1001/jamainternmed.  2020.0618. PMID: 32250383; PMCID: PMC7136858.
4. Role of Biomarkers in Health Care. Jain KK. The Handbook of Biomarkers. 2010 Jan 20:115–88. doi: 10.1007/978-1-60761-685-6_5. PMCID: PMC7123449.

 

World Kidney Day – March 9, 2023 – raising awareness of the importance of our kidneys

Around 1 in 10 people suffer from chronic kidney disease (CKD), with more than 800 million individuals being affected worldwide. CKD is a progressive disease in which the kidneys gradually lose their function over time. If detected early, medication and changes in lifestyle may help to prevent or slow down CKD progression.

What are the risk factors for CKD?

Older age (+60 years), diabetes, high blood pressure, heart disease, obesity and some medications are some known risk factors for kidney disease.

How can we keep our kidneys healthy?

Nutrition, exercise, sufficient fluid intake are only some examples on how to keep our kidneys healthy. Valuable information can be found on the following websites :

“World Kidney Day”

“The International Society of Nephrology – ISN”

“International Federation of Kidney Foundations – IFKF”

BIOMEDICA – Biomarker ELISA kits for clinical research in kidney disease

check out our Brochure – Biomarkers in Clinical Nephrology

ROBUST & RELIABLE ASSAYS

• Fully validated according to international quality guidelines
• Widely cited in over 1500 publications

FGF23 (Fibroblast growth factor 23) Vanin-1, Endostatin, Sclerostin, Osteoprotegerin, Angiopoietin-2, IL-6, VEGF

Biomedica Quality ELISA kits

LITERATURE

Plant-based diets for prevention and management of chronic kidney disease. Joshi S, Hashmi S, Shah S, Kalantar-Zadeh K. Curr Opin Nephrol Hypertens. 2020 Jan;29(1):16-21. doi: 10.1097/MNH.0000000000000574. PMID: 31725014. 

Abstract

Purpose of review: Plant-based diets have been used with growing popularity for the treatment of a wide range of lifestyle-related diseases, including diabetes, hypertension, and obesity. With the reinvigoration of the dietary management of chronic kidney disease (CKD) and the use of low protein diets for secondary prevention of CKD to delay or prevent dialysis therapy, there is an increasing interest in the potential role of plant-based diets for these patients.

Recent findings: Recently, a body of evidence related to the role of plant-based diets in preventing CKD has reemerged. Several observational studies have shown that red and processed meat have been associated with increased risk of CKD as well as faster progression in those with preexisting CKD. In several substitution analyses, replacement of one serving of red and/or processed meat has been linked with sizable reductions in CKD risk. Although limited, experimental trials for the treatment of metabolic acidosis in CKD with fruits and vegetables show outcomes comparable to oral bicarbonate. The use of plant-based diets in CKD may have other benefits in the areas of hypertension, weight, hyperphosphatemia, reductions in hyperfiltration, and, possibly, mortality. The risk of potassium overload from plant-based diets appears overstated, mostly opinion-based, and not supported by the evidence. Plant-based diets are generally well tolerated and provide adequate protein intake, including essential amino acids as long as the diet is correctly implemented.

Summary: Plant-based diets should be recommended for both primary and secondary prevention of CKD. Concerns of hyperkalemia and protein inadequacy related to plant-based diets may be outdated and unsupported by the current body of literature. Healthcare providers in general medicine and nephrology can consider plant-based diets as an important tool for prevention and management of CKD.

Exercise and Kidney Disease Prevention: Walk This Way. Seliger S, Weiner DE. Am J Kidney Dis. 2022 Oct;80(4):552-554. doi: 10.1053/j.ajkd.2022.07.001. Epub 2022 Jul 21. PMID: 35872228.

 

Raising awareness, screening and prevention of chronic kidney disease: It takes more than a village. Hsiao LL. Nephrology (Carlton). 2018 Oct;23 Suppl 4:107-111. doi: 10.1111/nep.13459. PMID: 30298651.

Abstract

Chronic kidney disease (CKD) is a major public health problem worldwide. Its prevalence and incidence are increasing, particularly among the ethnic minority populations. Diabetes, hypertension and obesity have been the three major aetiologies for CKD in all developed countries. While diabetes and hypertension remain the major causes of CKD in developing countries, environmental pollution, pesticides, water, analgesic abuse and herbal medications are common causes in these regions. Rapid urbanization and globalization are thought to be the contributing factors to rising prevalence and incidents of CKD. Despite the rising prevalence of CKD, disease awareness remains profoundly low. Worldwide, only 6% of the general population and 10% of the high-risk population are aware of their CKD statuses. Health screenings have been shown to be effective in improving the incidence of ESRD. However, currently there is no effective tool to assess and evaluate the awareness objectively.

Microbiome-metabolome reveals the contribution of gut-kidney axis on kidney disease. Chen YY, Chen DQ, Chen L, Liu JR, Vaziri ND, Guo Y, Zhao YY. J Transl Med. 2019 Jan 3;17(1):5. doi: 10.1186/s12967-018-1756-4. PMID: 30602367; PMCID: PMC6317198.

 

Acute kidney injury (AKI) – also called acute renal failure –  occurs when the kidneys suddenly fail to function due to an injury medication, or an infection. It happens within a few hours or days and is usually accompanied with other medical conditions e.g. blood vessel blockage, diabetes, heart failure, kidney and liver disease, hospitalization (ICU) and other.

FGF23 in Acute Kidney Injury

Fibroblast growth factor 23 (FGF23) is a bone derived hormone that regulates renal phosphate excretion in the kidney. In kidney disease, when the function of the kidney declines, serum phosphate levels rise which subsequently leads to the secretion of FGF-23. High phosphate levels are also common in patients with acute kidney injury (AKI) (1).

FGF23 marker of adverse outcomes in acute kidney injury 

 Fibroblast growth factor 23 (FGF23) levels rise rapidly with acute kidney injury and are associated with the requirement for renal replacement therapy (1-4). FGF-23 levels also have prognostic utility as shown in a large study in over 1500 patients with AKI (3).

Most studies measuring elevated FGF23 levels in AKI patients are performed using FGF23 (C-terminal) assays, which detect intact and C-terminal FGF23. Although the results mirror an increase of FGF23 production they may not necessarily reflect the bioactivity of FGF23 (5).

Quantifying circulating FGF23 levels

Circulating FGF23 levels include the bioactive intact hormone (iFGF23) and the inactive N-terminal and C-terminal fragments. FGF23 can be measured with two commercially available assay types (6). The C-terminal FGF23 assay captures both intact and the c-terminal fragments of FGF23, which result after cleavage. The intact FGF23 assay utilizes antibodies that are located in the N-terminal and in the C-terminal region of the FGF23 hormone, thus capturing only biologically active intact FGF23.

Learn more:  FGF23 – an Overview

BIOMEDICA offers quality FGF23 assays for serum & plasma samples

 

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

FGF23 intact ELISA, cat no. BI-20700

 

• developed & manufactured by Biomedica , Austria
• fully validated
• numerous top journal citations

 

Literature

1. Fibroblast Growth Factor 23 and Klotho in AKI. Christov M, Neyra JA, Gupta S, Leaf DE. Semin Nephrol. 2019 Jan;39(1):57-75. doi: 10.1016/j.semnephrol.2018.10.005. PMID: 30606408. 
2. Fibroblast Growth Factor 23 Regulation and Acute Kidney Injury. Zhou W, Simic P, Rhee EP. Nephron. 2022;146(3):239-242. doi: 10.1159/000517734. Epub 2021 Jul 20. PMID: 34284404; PMCID: PMC8770696. 
3. Fibroblast growth factor 23 associates with death in critically ill patients. Leaf DE, Siew ED, Eisenga MF, Singh K, Mc Causland FR, Srivastava A, Ikizler TA, Ware LB, Ginde AA, Kellum JA, Palevsky PM, Wolf M, Waikar SS, Am Clin J Am Soc Nephrol. 2018. 13(4):531–41. 
4. FGF-23 levels in patients with AKI and risk of adverse outcomes. Leaf DE, Wolf M, Waikar SS, Chase H, Christov M, Cremers S, Stern L. Clin J Am Soc Nephrol. 2012. 7(8):1217-23. 
5. Fibroblast Growth Factor 23 and αKlotho in Acute Kidney Injury: Current Status in Diagnostic and Therapeutic Applications. Neyra JA, Hu MC, Moe OW. Nephron. 2020;144(12):665-672. doi: 10.1159/000509856. Epub 2020 Aug 25. PMID: 32841947; PMCID: PMC7708396. 
6. The Measurement and Interpretation of Fibroblast Growth Factor 23 (FGF23) Concentrations. Heijboer AC, Cavalier E. Calcif Tissue Int. 2023. 112(2):258-270.

 

 

NEW – Rat NT-proBNP ELISA assay 

quantitative, reliable and robust for translational research and drug discovery.

Rat NT-proBNP ELISA assay │ BI-1204R

features & benefits

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

 

NT-proANP ELISA assay │BI-20892

 features & benefits

• 10 µl / well, serum or plasma
• widely cited as cardiovascular safety biomarker in rats

 

“cross-laboratory comparison study detecting serum NT-proANP in rats with the Biomedica NT-proANP ELISA to be technically adequate with acceptable intra- and inter-assay and inter-laboratory precision and accuracy (Vinken et al., 2016).”

All assays are developed and manufactured by Biomedica! 

NT-proBNP & NT-proANP in drug-induced cardiac hypertrophy

The cardiac biomarkers NT-proBNP and NT-proANP are predictive of left ventricular hypertrophy (LVH) and systolic dysfunction in humans (1, 2).

In preclinical settings they have successfully been used to detect cardiotoxicity (3). NT-proBNP and NT-proANP have shown to be useful tools that help to improve the detection of cardiovascular injury early in drug development (4).

REFERENCES

1. Circulating N-terminal pro-atrial natriuretic peptide is an independent predictor of left ventricular hypertrophy in the general population. The Tromsø Study. Schirmer H, Omland T. Eur Heart J. 1999 May;20(10):755-63. doi: 10.1053/euhj.1998.1396. Erratum in: Eur Heart J 1999 Oct;20(19):1439. PMID: 10329067.
2.  An association between N-terminal pro-brain natriuretic protein level and risk of left ventricular hypertrophy in patients without heart failure. Huang L, Huang L, Yu J, Wu X, Zhao J Exp Ther Med. 2020 May;19(5):3259-3266. doi: 10.3892/etm.2020.8598. Epub 2020 Mar 12. PMID: 32266021; PMCID: PMC7132238.
3.  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, Manfredi TG, Agostinucci K, Engle SK, Powe J, King NM, Rodriguez LA, Gropp KE, Gallacher M, Vetter FJ, More V, Shimpi P, Serra D, Colton HM Toxicol Pathol. 2017 Feb;45(2):344-352. doi: 10.1177/0192623316634231. Epub 2016 Jul 11. PMID: 27102652.
4.  Integrated approach to early detection of cardiovascular toxicity induced by a ghrelin receptor agonist. Stokes AH, Falls JG, Yoon L, Cariello N, Faiola B, Colton HM, Jordan HL, Berridge BR. Int J Toxicol. 2015 Mar-Apr;34(2):151-61. doi: 10.1177/1091581815573029. Epub 2015 Feb 26. PMID: 25722321.

 

Valvular heart disease is a leading cause of cardiovascular morbidity. Transcatheter aortic valve replacement (TAVR) is a minimally invasive procedure that replaces the aortic valve in patients with aortic stenosis.  Mineral homeostasis, the process that regulates the body’s levels of calcium and phosphorus, plays an important role in the progression of cardiovascular diseases, including calcific aortic valve stenosis. FGF23 is a bone-derived phosphotropic hormone that regulates phosphate and vitamin D metabolism. FGF23 has been shown to be an independent risk factor for CV morbidity and mortality.

FGF23 is a novel risk factor for patients undergoing TAVR

A study by Mirna et al. evaluated the predictive potential of both C-terminal FGF23 (cFGF23) and intact FGF23 (iFGF23) for adverse outcomes in patients undergoing transcatheter aortic valve replacement (TAVR) with regard to renal function. Patients were followed up at 12 months.

The authors demonstrated:

• significant association of high cFGF23 levels with mortality in patients with an estimated glomerular filtration rate (eGFR) ≥45 ml/min/1.73m².
• patients with cFGF23 levels above the cut-off of 6.82 pmol/l had a worse 1-year-mortality
• cFGF23 could be a novel risk factor for patients undergoing TAVR with eGFR ≥45ml/min/1.73m².

 

FGF23 (C-terminal) and FGF23 intact were both measured with an ELISA assay from BIOMEDICA.

 

Assay Highlights

• full validation package
• for serum and plasma
• +50 international references

 

LITERATURE

Serum levels of C-terminal FGF23 (cFGF23) are associated with 1-year-mortality in patients undergoing transcatheter aortic valve replacement (TAVR).

Mirna M, Lauten A, Jirak P, Rezar R, Wernly B, Paar V, Felder TK, Hoppe UC, Motloch LJ, Jung C, Alushi B, Lichtenauer M, Salmhofer H. Eur J Intern Med. 2021. 85:98-107. doi: 10.1016/j.ejim.2020.09.022. Epub 2020 Nov 13. PMID: 33191056.

Abstract

Introduction: Serum levels of FGF23 have been associated with adverse outcomes in cardiovascular diseases in patients with and without impaired renal function. Hence, this study aimed to explore the prognostic relevance of intact FGF23 (iFGF23) and its derivate C-terminal FGF23 (cFGF23) in patients undergoing transcatheter aortic valve replacement (TAVR) with regard to renal function.

Methods: A total of 274 patients undergoing transfemoral TAVR were enrolled in this study. Blood samples were obtained preinterventionally and analyzed for iFGF23 and cFGF23 by means of enzyme linked immunosorbent assay (ELISA). Follow-up was obtained for 12 months.

Results: Serum levels of cFGF23 and iFGF23 both correlated positively with serum creatinine and inversely with estimated glomerular filtration rate (eGFR). Cox regression analysis revealed a significant association of cFGF23 with 1-year-mortality in patients with eGFR ≥45ml/min/1.73m², but not in patients with an eGFR <45ml/min/1.73m². A cut-off was calculated for cFGF23 (6.82 pmol/l) and patients with eGFR ≥45ml/min/1.73m² were retrospectively divided into two groups (above/below cut-off). Patients above the cut-off had a significantly worse 1-year-mortality than patients below the cut-off (33.3% vs. 19.6%; OR 2.05 (95%CI 1.03-4.07), p= 0.038). The association of cFGF23 with 1-year-mortality in patients with eGFR ≥45ml/min/1.73m² remained statistically significant even after correction for possible confounders in a multivariate Cox regression analysis.

Conclusion: cFGF23 could be an individual risk factor for mortality in patients undergoing TAVR with an eGFR ≥45ml/min/1.73m².

 

 

 

The natriuretic peptides NT-proBNP & NT-proANP

Cardiac Biomarkers – kits for clinical & preclinical use

NT-proBNP is released in the heart in response to cardiac wall stretch or pressure overload.   It is considered as the gold standard in heart failure and is useful for both diagnosis and prognosis (1).

NT-proANP is synthesized and stored in atrial myocytes in response to increased intra-atrial  pressure. Elevated circulating levels of NT-proANP are associated with heart failure resulting from increased mechanical stress in the heart during hypertension. In population-based studies, elevated plasma NT-pro-ANP levels predicted the risk of cardiovascular events and death ( 2).

 

Cardiac Biomarkers – kits for clinical & preclinical use

Natriuretic peptides in pre-clinical models as cardiac toxicity biomarkers

Biomarkers are also a valuable tool to explore drug effects in pre-clinical models. The natriuretic peptide NT-proANP has successfully been validated as a cardiovascular safety biomarker in rodents (3). An additional cardiac toxicity biomarker in drug development is NT-proBNP supporting the use of natriuretic peptides to investigate drug-induced cardiac hypertrophy (4, 5).

Biomedica offers a range of top quality ELISA kits for your clinical & preclinical research

 

• NT-proBNP – human (CE-marked in EU) (#SK-1204)
• NT-proBNP – rat (new!)  (#BI-1204R)
• NT-proANP – human, rodent (#BI-20892) citations rat/mouse

 

 

• widely cited in over 400 publications
• kit validation follows international quality guidelines

 

LITERATURE 

1. NT-proBNP: The Gold Standard Biomarker in Heart Failure. McKie PM, Burnett JC Jr. J Am Coll Cardiol. 2016; 6;68(22):2437-2439. 
2. Plasma natriuretic peptide levels and the risk of cardiovascular events and death. Wang TJ, Larson MG, Levy D, Benjamin EJ, Leip EP, Omland T, Wolf PA, Vasan RS. N Engl J Med. 2004; 12;350(7):655-63. 
3. 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. Pharmacol Toxicol Methods. 2016; 77:58-65.  
4. Evaluation of Cardiac Toxicity Biomarkers in Rats from Different Laboratories. Kim K, Chini N, Fairchild DG, Engle SK, Reagan WJ, Summers SD, Mirsalis JC – Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium. Toxicol Pathol. 2016; 44(8):1072-1083. 
5. Serum Natriuretic Peptides as Differential Biomarkers Allowing for the Distinction between Physiologic and Pathologic Left Ventricular Hypertrophy. Dunn ME, Manfredi TG, Agostinucci K, Engle SK, Powe J, King NM, Rodriguez LA, Gropp KE, Gallacher M, Vetter FJ, More V, Shimpi P, Serra D, Colton HM – Cardiac Hypertrophy Working Group of the Predictive Safety Testing Consortium. Toxicol Pathol. 2017; 45(2):344-352.

 

For use in preclinical models – cardiotoxicity biomarkers

Rat NT-proBNP ELISA , cat. no. BI-1204R

NT-proANP ELISA independently validated for rat, cat. no. BI-20892

February is “Heart Month” raising awareness about heart disease. Biomarker research has revolutionized on how heart diseases are diagnosed and treated. Ongoing research on newer exploratory protein biomarkers may shed light into the complex mechanisms that drive the disease process.

Novel Biomarkers for Heart Disease

BIG ENDOTHELIN-1, ENDOSTATIN, FGF23, LRG

 

Big Endothelin-1 (Big ET-1) is a vasoconstrictor peptide and is the precursor of Endothelin-1 (ET-1) the biologically active form. Elevated concentrations provide an independent indicator of heart failure in congestive heart disease (1). Big ET-1 has a longer half-life than ET-1 and circulates in equimolar amounts as ET-1, making it a more reliable biomarker.

Endostatin is a degradation product of collagen XVIII and is an extracellular matrix protein. Endostatin plays an essential role in the pathogenesis of chronic kidney and heart diseases (2). A study in two community-based cohorts of elderly showed an association of high serum endostatin levels with left ventricular dysfunction and an increased heart failure risk (3).  

Fibroblast growth factor-23 (FGF23) is a hormone that regulates phosphate metabolism. FGF23 has been suggested as a novel candidate biomarker of cardiovascular risk. High circulating FGF23 levels have been associated with adverse cardiovascular outcomes such as heart failure and arrhythmias (4).

Leucine-riche alpha-2-glyoprotein (LRG) is a secreted protein that plays an important role in pathogenic ocular neovascularization. LRG is also involved in multiple conditions including cardiovascular disease, diabetes, inflammatory disorders, and cancer. A recent study demonstrated that LRG accurately identify patients with heart failure stronger than BNP, independent of age, sex, and creatinine (5, 6).

NOVEL BIOMARKERS FOR HEART DISEASE

Biomedica offers a range of top quality ELISA kits for your clinical & preclinical research.

-Big Endothelin-1 (#BI-20082H)

-Endostatin (#BI-20742)

-Endostatin mouse/rat (#BI-20742MR)

-FGF23 intact (#BI-20700)

-FGF23 c-terminal (#BI-20702)

-LRG (#BI-LRG)

 

widely cited in over 130 publications kit validation follows international quality guidelines

LITERATURE

1. Plasma concentration of big endothelin-1 and its relation with plasma NT-proBNP and ventricular function in heart failure patients. Rivera M et al., Rev Esp Cardiol. 2005;  58(3):278-84.
2. Endostatin in Renal and Cardiovascular Diseases. Li M et al., Kidney Dis. 2021; 9;7(6):468-481.
3. Circulating endostatin and the incidence of heart failure. Ruge T et al., 2018; 52(5):244-249.
4. Fibroblast Growth Factor 23 and Long-Term Cardiac Function: The Multi-Ethnic Study of Atherosclerosis. Patel RB et al., Cardiovasc Imaging. 2020; 13(11):e011925.
5. Proteomic analysis of coronary sinus serum reveals leucine-rich α2-glycoprotein as a novel biomarker of ventricular dysfunction and heart failure. Watson CJ et al., Circ Heart Fail. 2011; 4(2):188-97.
6. Serum biomarker discovery related to pathogenesis in acute coronary syndrome by proteomic approach  Shin M et al.,Biosci Rep. 2021; 25;41(6):BSR20210344.

Widely cited non-radioactive EZ4U – Cell Proliferation & Cytotoxicity Assay – single step incubation for use on living cells

The Biomedica EZ4U cell proliferation and cytotoxicity assay  was highlighted in a recent study investigating the cytotoxicity of drug combinations tested against different pancreatic cell lines.  Learn more: Cytotoxicity of combinations of the pan-KRAS SOS1 inhibitor BAY-293 against pancreatic cancer cell lines.

Pancreatic ductal adenocarcinomas (PDACs), the most prevalent pancreatic cancer, is highly aggressive with a 5-year overall survival rate of less than 8%.  Late detection, it´s metastatic spread and the resistance to chemotherapy are among some of the factors responsible for poor patient outcome. The PDAC incidence rate is increasing and is particularly related to the general aging of our society. Furthermore, life style habits that include abuse of alcohol and tobacco as well as obesity and type 2 diabetes increases the risk for various types of cancer including PDAC.

EZ4U – Cell Proliferation & 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 230 publications

BROCHURE – EZ4U cell proliferation and cytotoxicity assay

 

FURTHER READING

Pancreatic ductal adenocarcinoma: Treatment hurdles, tumor microenvironment and immunotherapy.

World J Gastrointest Oncol. Sarantis P, Koustas E, Papadimitropoulou A, Papavassiliou AG, Karamouzis MV. 2020 Feb 15;12(2):173-181. doi: 10.4251/wjgo.v12.i2.173. PMID: 32104548; PMCID: PMC7031151.

 

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal diseases, with an average 5-year survival rate of less than 10%. Unfortunately, the majority of patients have unresectable, locally advanced, or metastatic disease at the time of diagnosis. Moreover, traditional treatments such as chemotherapy, surgery, and radiation have not been shown to significantly improve survival. Recently, there has been a swift increase in cancer treatments that incorporate immunotherapy-based strategies to target all the stepwise events required for tumor initiation and progression. The results in melanoma, non-small-cell lung cancer and renal cell carcinoma are very encouraging. Unfortunately, the application of checkpoint inhibitors, including anti-CTLA4, anti-PD-1, and anti-PD-L1 antibodies, in pancreatic cancer has been disappointing. Many studies have revealed that the PDAC microenvironment supports tumor growth, promotes metastasis and consists of a physical barrier to drug delivery. Combination therapies hold great promise for enhancing immune responses to achieve a better therapeutic effect. In this review, we provide an outline of why pancreatic cancer is so lethal and of the treatment hurdles that exist. Particular emphasis is given to the role of the tumor microenvironment, and some of the latest and most promising studies on immunotherapy in PDAC are also presented.

 

Pancreatic ductal adenocarcinoma: biological hallmarks, current status, and future perspectives of combined modality treatment approaches.

Orth M, Metzger P, Gerum S, Mayerle J, Schneider G, Belka C, Schnurr M, Lauber K. Radiat Oncol. 2019 Aug 8;14(1):141. doi: 10.1186/s13014-019-1345-6. PMID: 31395068; PMCID: PMC6688256.

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a highly devastating disease with poor prognosis and rising incidence. Late detection and a particularly aggressive biology are the major challenges which determine therapeutic failure. In this review, we present the current status and the recent advances in PDAC treatment together with the biological and immunological hallmarks of this cancer entity. On this basis, we discuss new concepts combining distinct treatment modalities in order to improve therapeutic efficacy and clinical outcome – with a specific focus on protocols involving radio(chemo)therapeutic approaches.

 

Pancreatic ductal adenocarcinomas (PDACs), the most prevalent pancreatic cancer, is highly aggressive with a 5-year overall survival rate of less than 8%.  Late detection, it´s metastatic spread and the resistance to chemotherapy are among some of the factors responsible for poor patient outcome. The PDAC incidence rate is increasing and is particularly related to the general aging of our society. Furthermore, life stye habits that include abuse of alcohol and tobacco as well as obesity and type 2 diabetes increases the risk for various types of cancer including PDAC.

 

Do you need support in meeting your research goals? We can help! We offer custom analytical service measurements for ELISA Kits, Luminex Assays, or microRNA Analysis .

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Custom Analytical Service Measurements for ELISA, Luminex, microRNA

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RELATED PUBLICATIONS

Comprehensive Characterization of Platelet-Enriched MicroRNAs as Biomarkers of Platelet Activation.

Krammer TL, Zeibig S, Schrottmaier WC, Pirabe A, Goebel S, Diendorfer AB, Holthoff HP, Assinger A, Hackl M. Cells. 2022 Apr 7;11(8):1254. doi: 10.3390/cells11081254. PMID: 35455934; PMCID: PMC9030873.

Abstract

Dysregulation of platelet function is causally connected to thrombus formation and cardiovascular diseases. Therefore, assessing platelet reactivity is crucial. However, current platelet function tests come with pitfalls, limiting clinical use. Plasma miRNA signatures have been suggested as novel biomarkers for predicting/diagnosing cardiovascular diseases and monitoring antiplatelet therapy. Here, we provide results from a comprehensive study on the feasibility of using circulatory platelet miRNAs as surrogate markers of platelet activation. We performed small RNA-Seq on different blood cell types to confirm known and identify novel platelet-enriched miRNAs and validated a panel of 16 miRNAs using RT-qPCR. To identify the main carrier of these blood-based platelet miRNAs, we enriched and analyzed distinct microvesicle populations. Platelets were stimulated with GPVI and P2Y12 agonists in vitro to monitor the release of the selected miRNAs following activation. Finally, the miRNA panel was also measured in plasma from mice undergoing the Folts intervention (recurrent thrombus formation in the carotid artery). Applying an unbiased bioinformatics-supported workflow to our NGS data, we were able to confirm a panel of previously established miRNA biomarker candidates and identify three new candidates (i.e., miR-199a-3p, miR-151a-5p, and miR-148b-3p). Basal levels of platelet-derived miRNAs in plasma were mainly complexed with proteins, not extracellular vesicles. We show that changes in miRNA levels due to platelet activation are detectable using RT-qPCR. In addition, we highlight limitations of studying the in vitro release of miRNAs from platelets. In vivo thrombosis resulted in significant elevations of platelet-derived miRNA levels in mice. In conclusion, we provide in-depth evidence that activated platelets release miRNAs, resulting in measurable changes in circulatory miRNA levels, rendering them promising biomarker candidates.

Multiplex Soluble Biomarker Analysis from Pleural Effusion.

Biomolecules. Javadi J, Dobra K, Hjerpe A. 2020 Jul 28;10(8):1113. doi: 10.3390/biom10081113. PMID: 32731396; PMCID: PMC7464384.

Abstract

Malignant pleural mesothelioma (MPM) is a highly aggressive and therapy resistant pleural malignancy that is caused by asbestos exposure. MPM is associated with poor prognosis and a short patient survival. The survival time is strongly influenced by the subtype of the tumor. Dyspnea and accumulation of pleural effusion in the pleural cavity are common symptoms of MPM. The diagnostic distinction from other malignancies and reactive conditions is done using histopathology or cytopathology, always supported by immunohistochemistry, and sometimes also by analyses of soluble biomarkers in effusion supernatant. We evaluated the soluble angiogenesis related molecules as possible prognostic and diagnostic biomarkers for MPM by Luminex multiplex assay. Pleural effusion from 42 patients with malignant pleural mesothelioma (MPM), 36 patients with adenocarcinoma (AD) and 40 benign (BE) effusions were analyzed for 10 different analytes that, in previous studies, were associated with angiogenesis, consisting of Angiopoietin-1, HGF, MMP-7, Osteopontin, TIMP-1, Galectin, Mesothelin, NRG1-b1, Syndecan-1 (SDC-1) and VEGF by a Human Premixed Multi-Analyte Luminex kit. We found that shed SDC-1 and MMP-7 levels were significantly lower, whereas Mesothelin and Galectin-1 levels were significantly higher in malignant mesothelioma effusions, compared to adenocarcinoma. Galectin-1, HGF, Mesothelin, MMP-7, Osteopontin, shed SDC-1, NRG1-β1, VEGF and TIMP-1 were significantly higher in malignant pleural mesothelioma effusions compared to benign samples. Moreover, there is a negative correlation between Mesothelin and shed SDC-1 and positive correlation between VEGF, Angiopoietin-1 and shed SDC-1 level in the pleural effusion from malignant cases. Shed SDC-1 and VEGF have a prognostic value in malignant mesothelioma patients. Collectively, our data suggest that MMP-7, shed SDC-1, Mesothelin and Galectin-1 can be diagnostic and VEGF and SDC-1 prognostic markers in MPM patients. Additionally, Galectin-1, HGF, Mesothelin, MMP-7, Osteopontin, shed SDC-1 and TIMP-1 can be diagnostic for malignant cases.

Biomarkers & Bone Health – ELISA Kits for Clinical Research

Bone remodeling is a continuous process that removes bone and replaces it with newly synthesized bone. This bone turnover process preserves the mechanical function of the human skeleton.

Bone turnover biomarkers, e.g. markers of bone formation and bone resorption, have been used during the last decade to monitor bone diseases and to monitor their treatment.

Many of these markers are secreted by osteoblasts and osteoclasts and include regulators of bone turnover e.g. receptor activator of NF-kB ligand (RANKL) and osteoprotegerin (OPG).  

Though RANKL and OPG  play an integral role in bone turnover, they do not reflect the activity of osteocytes, the most abundant cell type in the bone.

Osteocytes are cells that regulate bone remodeling. They secrete proteins – bone regulation markers – that include Sclerostin (SOST), Dickkopf-1 (DKK-1), and Fibroblast growth factor  (FGF23). These markers reflect the osteocyte activity.

The above listed biomarkers circulate and can be measured in serum and plasma allowing the investigation of complex interactions between the bone and their relationship with other organs.

BIOMEDICA ELISA KITS – Biomarkers & Bone Health

check out our Bone Biomarker Brochure

ELISA 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

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

RELATED PRODUCTS

Osteoprotegerin (OPG) ELISA , soluble RANKL ELISA, Periostin ELISA, DKK-1 ELISA ,  Sclerostin ELISA

FGF23 ELISA , IL-6 ELISA  ,  VEGF ELISA ,  Angiopoietin-2 ELISA

MICRORNA  Analysis 

osteomiR^®– bone miRNA biomarkers highlights:

• Novel minimally invasive biomarkers to detect high imminent fracture-risk in osteoporosis
• 19 individual bone-related biomarkers with distinct information content

RELATED PUBLICATIONS

Novel biological markers of bone: from bone metabolism to bone physiology. Rheumatology (Oxford). Chapurlat RD, Confavreux CB. 2016; 55(10):1714-25.

Abstract

Biochemical markers of bone turnover have been used for decades in the management of bone diseases, to assess the prognosis of these conditions and to monitor treatments. The new markers, however, also reflect specific physiological mechanisms in the bone or other organs. Periostin may be more specific to the periosteum; cathepsin K is an osteoclastic enzyme that may be involved in the cardiovascular system and joints; Dickkopf-1 is involved in bone formation and vascular calcification; sclerostin is a major regulator of bone formation in response to mechanical loading and may also play a role in chronic kidney disease bone and mineral disorder; sphingosine-1-phosphate is a lipid mediator interacting with bone resorption. Some of the bone markers are in fact hormones produced by the bone that affect various physiological and pathological functions in other organs. Thus, osteocalcin is produced by osteoblasts and participates in the regulation of insulin sensitivity and fertility in men. Fibroblast growth factor 23 is produced by osteocytes to regulate phosphorus and 1,25(OH)2D3, but it also plays a major role in the adverse consequences of declining renal function, in particular with respect to the myocardium. Micro RNAs are single-stranded RNAs that regulate several pathways, including the development timing, organogenesis, cell apoptosis, proliferation and differentiation. Their serum concentration may reflect the links between bone physiology and certain conditions in other organs, for example, the cardiovascular system.

Bones have many important biological functions. Bone biomarkers have gained attention in clinical research for the assessment of bone-related diseases. Some of the biomarker proteins have been found to represent useful targets for therapeutic antibodies.

Biomarkers in Bone Biology

Function of the human skeleton

The human skeletal system gives the body it´s structure and helps to protect and support the internal organs. It forms a part of the muscular-skeletal systems that helps the body to move. Throughout our lifetime, the human skeleton undergoes constant remodeling. This dynamic process, degrading bone and replacing it with new tissue maintains bone mass. The continuous cycle of bone resorption and bone growth is also known as bone metabolism.

Bone remodeling

Bone remodeling is a tightly regulated process performed by hormones, cell-signaling molecules, and bone cells. These specific bone cells are osteoclasts, osteoblasts, and osteocytes. The cells are in constant communication with each other through secreted factors, such as osteoprotegerin, RANKL, and sclerostin. These regulatory systems keep the bone remodeling balanced. Imbalances in bone metabolism can lead to bone diseases.

Role of RANKL, RANK, and OPG in bone biology

 Bones are broken down by osteoclasts and rebuilt by osteoblasts.

RANKL receptor activator is a mediator of bone resorption and OPG acts as a decoy receptor.

 

Osteoprotegerin (OPG) is produced by osteoblasts, cells that synthesize bone. OPG is a decoy receptor and binds to RANKL, antagonizing its binding to RANK.

RANKL (receptor activator of nuclear factor kappa-B ligand) is secreted by osteoblasts and binds to the RANK receptor on osteoclast precursor and mature osteoclast cells. RANKL stimulates bone resorption.

 

Role of Sclerostin, FGF23, DKK-1, and Periostin in bone biology

 

 

Biomarkers in Bone Biology

 

Bone cells have been reported to have endocrine functions that affect multiple organs. The most abundant cell type in the bone are osteocytes residing within the bone matrix and comprising 90% to 95% of the bone cells. Osteocytes play a significant role in the regulation of osteogenesis, releasing osteocyte-related biomarkers such as sclerostin (SOST), fibroblast growth factor 23 (FGF23), and Dickkopf-1 (DKK-1).

Sclerostin (SOST) is mainly produced by osteocytes and is considered as the major regulator of bone formation. More recently, Sclerostin has been shown to stimulate the osteocyte synthesis of fibroblast growth factor-23, potentially contributing to the regulation of phosphate homeostasis.

Fibroblast growth factor 23 (FGF23) is a hormone that is mainly secreted by osteocytes and osteoblasts. It regulates phosphate and vitamin D levels and functions as a central endocrine hormone regulating phosphate balance.

Dickkopf-1 (DKK-1) is an extracellular protein. DKK-1 plays a role in the regulation of bone metabolism, as it inhibits the differentiation of osteoblasts.

Periostin (POSTN) is an extracellular matrix protein that is preferentially expressed in the periosteum, a membrane covering the outer surface of bones which is responsible for growth.  Periostin has functions in osteology, tissue repair, oncology, cardiovascular and respiratory diseases, and in a variety of  inflammatory settings (e.g. asthma).

BIOMEDICA OFFERS HIGH QUALITY ELISA KITS FOR BONE BIOMARKERS 

 

widely cited in over 1100 publications

kit validations follow international quality guidelines

 

Sclerostin ELISA (cat.no. BI-20492)

OPG ELISA (cat.no. BI-20403)

RANKL ELISA (cat.no. BI-20462)

FGF23 intact ELISA (cat.no. BI-20700)

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

DKK-1 ELISA (cat.no. BI-20413)

PERIOSTIN ELISA (cat.no. BI-20433)

 

RELATED LITERATURE

A Review of the Potential Application of Osteocyte-Related Biomarkers, Fibroblast Growth Factor-23, Sclerostin, and Dickkopf-1 in Predicting Osteoporosis and Fractures. Ramli FF, Chin KY. Diagnostics (Basel). 2020 Mar 6;10(3):145. doi: 10.3390/diagnostics10030145. PMID: 32155909; PMCID: PMC7151094.

Sclerostin Directly Stimulates Osteocyte Synthesis of Fibroblast Growth Factor-23. Wijenayaka AR, Yang D, Ormsby RT, Bonewald LF, Atkins GJ. Calcif Tissue Int. 2021 Jul;109(1):66-76. doi: 10.1007/s00223-021-00823-6. Epub 2021 Feb 22. PMID: 33616712.

Biology of the RANKL-RANK-OPG System in Immunity, Bone, and Beyond. Front Immunol. Walsh MC, Choi Y.2014 Oct 20;5:511. doi: 10.3389/fimmu.2014.00511. PMID: 25368616; PMCID: PMC4202272.

The Osteocyte: New Insights. Robling AG, Bonewald LF. Annu Rev Physiol. 2020 Feb 10;82:485-506. doi: 10.1146/annurev-physiol-021119-034332. PMID: 32040934; PMCID: PMC8274561.

 

 

Sema4D induces cartilage destruction

Semaphorin 4D destroys cartilage: Semaphorin 4D (Sema 4D) has been identified as an inflammatory cytokine that promotes cartilage destruction. This finding was recently published by Murakami T and colleagues. In a mouse model of inflammatory arthritis, the researchers demonstrated that Sema4D was increased in synovial fluid and loss of Sema4D protected against cartilage degeneration. Click here to learn more.

 

 

What is cartilage?

Cartilage is a type of strong and flexible tissue that is found throughout the body. It is a connective tissue that exists in our joints, bones, spine, ears and nose. It protects our joints and our bones, absorbing impact, reducing friction and helping our joints to move smoothly.

 If cartilage is degraded, movements will cause friction and pain, leading to swelling and stiffness. Cartilage degradation can be caused through direct injury, osteoarthritis, and aging.  

Semaphorin 4D destroys cartilage

Semaphorin 4D induces articular cartilage destruction and inflammation in joints by transcriptionally reprogramming chondrocytes.

Murakami T, Takahata Y, Hata K, Ebina K, Hirose K, Ruengsinpinya L, Nakaminami Y, Etani Y, Kobayashi S, Maruyama T, Nakano H, Kaneko T, Toyosawa S, Asahara H, Nishimura R. Sci Signal. 2022 Nov;15(758):eabl5304. doi: 10.1126/scisignal.abl5304. Epub 2022 Nov 1. PMID: 36318619.

Abstract

Proinflammatory cytokines play critical roles in the pathogenesis of joint diseases. Using a mass spectrometry-based cloning approach, we identified Semaphorin 4D (Sema4D) as an inflammatory cytokine that directly promoted cartilage destruction. Sema4d-deficient mice showed less cartilage destruction than wild-type mice in a model of rheumatoid arthritis. Sema4D induced a proinflammatory response in mouse articular chondrocytes characterized by the induction of proteolytic enzymes that degrade cartilage, such as matrix metalloproteinases (MMPs) and aggrecanases. The activation of Mmp13 and Mmp3 expression in articular chondrocytes by Sema4D did not depend on RhoA, a GTPase that mediates Sema4D-induced cytoskeletal rearrangements. Instead, it required NF-κB signaling and Ras-MEK-Erk1/2 signaling downstream of the receptors Plexin-B2 and c-Met and depended on the transcription factors IκBζ and C/EBPδ. Genetic and pharmacological blockade of these Sema4D signaling pathways inhibited MMP induction in chondrocytes and cartilage destruction in femoral head organ culture. Our results reveal a mechanism by which Sema4D signaling promotes cartilage destruction.

SEMA4D ELISA kit highlights

First fully validated Sema4D assay (cat.no. BI-20405)

10 µl sample/well

Reference values provided

Developed & manufactured by Biomedica

RELATED PUBLICATIONS

Inhibition of Semaphorin 4D/Plexin-B1 signaling inhibits the subchondral bone loss in early-stage osteoarthritis of the temporomandibular joint. Zhang Z, Lu L, Ye T, Yu S, Zhang J, Zhang M, He F, Liu Q, Yang H, Feng J.Arch Oral Biol. 2022 Mar;135:105365. doi: 10.1016/j.archoralbio.2022.105365. Epub 2022 Feb 2. PMID: 35151027.

Abstract