Different conformational forms of serum carnosinase detected by a newly developed sandwich ELISA for the measurements of carnosinase concentrations

Amino Acids
Katja AdelmannSibylle J Hauske

Abstract

Serum carnosinase (CN-1) measurements are at present mainly performed by assessing enzyme activity. This method is time-consuming, not well suited for large series of samples and can be discordant to measurements of CN-1 protein concentrations. To overcome these limitations, we developed sandwich ELISA assays using different anti-CN-1 antibodies, i.e., ATLAS (polyclonal IgG) and RYSK173 (monoclonal IgG1). With the ATLAS-based assay, similar amounts of CN-1 were detected in serum and both EDTA and heparin plasma. The RYSKS173-based assay detected CN-1 in serum in all individuals at significantly lower concentrations compared to the ATLAS-based assay (range: 0.1-1.8 vs. 1-50 μg/ml, RYSK- vs. ATLAS-based, P<0.01). CN-1 detection with the RYSK-based assay was increased in EDTA plasma, albeit at significantly lower concentrations compared to ATLAS. In heparin plasma, CN-1 was also poorly detected with the RYSK-based assay. Addition of DTT to serum increased the detection of CN-1 in the RYSK-based assay almost to the levels found in the ATLAS-based assay. Both ELISA assays were highly reproducible (R: 0.99, P<0.01 and R: 0.93, P<0.01, for the RYSK- and ATLAS-based assays, respectively). Results of the ATLAS-based assay showed a posit...Continue Reading

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Citations

Jul 25, 2015·Amino Acids·Verena PetersEmile de Heer
Apr 5, 2016·Obesity·Barbora de CourtenBarbara Ukropcova
Oct 14, 2017·Journal of Inherited Metabolic Disease·Verena PetersClaus P Schmitt
Jan 11, 2014·Journal of Applied Physiology·Audrey BaguetWim Derave
Sep 22, 2015·American Journal of Physiology. Endocrinology and Metabolism·Sanne StegenWim Derave
Aug 18, 2020·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Jiedong QiuBenito A Yard
Mar 28, 2019·Current Medicinal Chemistry·Verena PetersClaus Peter Schmitt
Jul 31, 2021·Molecular Nutrition & Food Research·Jordi Mayneris-PerxachsBarbora de Courten

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