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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

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Volume 43, Issue 5


Two immunochemical assays to measure advanced glycation end-products in serum from dialysis patients

Xiaohong Zhang / Matthias Frischmann / Rose Kientsch-Engel / Katharina Steinmann / Helga Stopper / Toshimitsu Niwa / Monika Pischetsrieder
Published Online: 2011-09-21 | DOI: https://doi.org/10.1515/CCLM.2005.089


Advanced glycation end-products are uremic toxins that accumulate in the serum and tissues of patients with chronic renal failure. Here, we established two enzyme-linked immunosorbent assays (ELISAs) for N ε-carboxymethyllysine and imidazolone to analyze advanced glycation end-products in human serum. Both ELISAs detected advanced glycation end-products bound to human serum albumin in a dose-dependent way. Whereas the formation of imida-zolone was independent of the presence of oxygen, concentrations of N ε-carboxymethyllysine epitopes increased 20-fold under oxidative conditions. The N ε-carboxymethyllysine ELISA showed a similar response to free, peptide-bound and protein-bound N ε-carboxymethyllysine, whereas the imidazolone antibody showed slightly higher affinity toward peptide-bound compared to protein-bound imidazolone. In human serum, linear dilution ranges from 1:10 to 1:40 (N ε-carboxymethyllysine ELISA) and from 1:2 to 1:8 (imidazolone ELISA) were found. The recovery of N ε-carboxymethyllysine from serum was 101±10% and 94±12%, respectively, and 93±15% and 97±12% for imidazolone. The coefficients of variation for intra-assay variability were 0.26–2.7% (N ε-carboxymethyllysine) and 0.1–2.4% (imidazolone), and 8.3–13.4% (N ε-carboxymethyllysine) and 7.8–12.5% (imidazolone) for inter-assay variability. In serum samples from hemodialysis patients (n=20) and controls (n=20), an approximately two-fold increase was detected in the patient group (p<0.001). The combination of the N ε-carboxymethyllysine and imidazolone ELISAs is a valuable tool to measure serum concentrations of advanced glycation end-products for clinical studies.

Keywords: advanced glycation end-products; N ε-carboxymethyllysine (CML); enzyme-linked immunosorbent assay (ELISA); imidazolone; serum


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About the article

Corresponding author: Dr. Monika Pischetsrieder, Institute of Pharmacy and Food Chemistry, University of Erlangen-Nürnberg, Schuhstr. 19, 91052 Erlangen, Germany Phone: +49-9131-852-4102, Fax: +49-9131-852-2587, E-mail:

Received: 2004-11-23

Accepted: 2005-03-03

Published Online: 2011-09-21

Published in Print: 2005-05-01

Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 43, Issue 5, Pages 503–511, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2005.089.

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