Abstract
Estimation of advanced glycation end products (AGEs) by determining fluorescence is based on the use of a standard calibrator prepared by incubating bovine serum albumin (BSA) and glucose at 37°C for 60 days. In the present study we attempted to reduce the duration of incubation to 4 days by increasing the temperature to 50°C. It is noteworthy that incubation at 50°C resulted in the rapid production of an AGE‐BSA standard calibrator within 4 days. Aminoguanidine reduced the intensity of the glycation‐induced fluorescence, while the addition of lysine intensified the reaction, as shown by the calibrator incubated at 37°C. The protein carbonyl content was shown to increase in the rapidly‐formed standard calibrator. Thus we conclude that a simple increase in temperature and the addition of lysine (0.1M) can accelerate the process of glycation‐induced fluorescence. This calibrator can be used effectively in fluorescence assays of AGEs. J. Clin. Lab. Anal. 19:11–15, 2005. © 2005 Wiley‐Liss, Inc.
Keywords: diabetes, glycation, AGE‐BSA, lysine, fluorescence, advanced glycation end products
REFERENCES
- 1. Makita Z, Vlassara H, Cerami A, Bucala R. Immunochemical detection of advanced glycosylation end products in vivo. J Biol Chem 1992;267:5133–5138. [PubMed] [Google Scholar]
- 2. Vlassara H, Palace MR. Diabetes and advanced glycation endproducts. J Int Med 2002;251:87–101. [DOI] [PubMed] [Google Scholar]
- 3. Stitt AW, Jenkins AJ, Cooper ME. Advanced glycation end products and diabetic complications. Expert Opin Invest Drugs 2002;11:1–19. [DOI] [PubMed] [Google Scholar]
- 4. Rahbar S, Nadler JL. A new rapid method to detect inhibition of Amadori product generated by δ‐gluonlactone. Clin Chim Acta 1999;287:123–130. [DOI] [PubMed] [Google Scholar]
- 5. Wrobel K, Worbel K, Garay‐Sevilla E, Nava LE, Malacara JM. Novel analytical approach to monitoring advanced glycosylation end product in human serum with on‐line spectrophotometric and spectrofluorometric detection in a flow system. Clin Chem 1997;43:1563–1569. [PubMed] [Google Scholar]
- 6. Monnier VM, Kohn RR, Cerami A. Accelerated age‐related browning of human collagen in diabetes mellitus. Proc Natl Acad Sci USA 1984;81:583–587. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7. Xu X, Li Z, Luo D, Huang Y, et al. Exogenous advanced glycosylation end products induce diabetes like vascular dysfunction in normal rats: a factor in diabetic retinopathy. Graefe's Arch Clin Exp Ophthalmol 2003;41:56–62. [DOI] [PubMed] [Google Scholar]
- 8. Semdsrod B, Melkko J, Araki N, Sano H, Horiuchi S. Advanced glycation end products are eliminated by scavenger‐receptor‐mediated endocytosis in hepatic sinusoidal Kupffer and endothelial cells. Biochem J 1997;322:567–573. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9. Kilhovd BK, Thorsby P, Berg TJ, Hanssen KF, Birkeland KI. Serum levels of advanced glycation end products are increased in patients with type 2 diabetes and coronary heart disease. Diabetes Care 1999;22:1543–1548. [DOI] [PubMed] [Google Scholar]
- 10. Uchida K, Kanemastu K, Sakai K, et al. Protein bound acrolein: potential markers for oxidative stress. Proc Natl Acad Sci USA 1998;95:4882–4887. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11. Sharma SD, Pandey BN, Mishra KP, Sivakami S. Amadori product and age formation during nonenzymatic glycosylation of bovine serum albumin in vitro. J Biochem Mol Biol Biophys 2002;6:233–242. [DOI] [PubMed] [Google Scholar]
- 12. Miyata S, Monnier V. Immunochemical detection of advanced glycosylation end products in diabetic tissues using monoclonal antibody to pyrraline. J Clin Invest 1992;89:1102–1112. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13. Traverso N, Menini S, Cottalasso D, Odetti P, Marinari UM, Pronzato MA. Mutual interaction between glycation and oxidation during non‐enzymatic protein modification. Biochim Biophys Acta 1997;1336:409–418. [DOI] [PubMed] [Google Scholar]
- 14. Vlassara H, Striker LJ, Teichberg S, Fuh H, Li YM, Steffes M. Advanced glycation end products induce glomerular sclerosis and albuminuria in normal rats. Proc Natl Acad Sci USA 1994;91:11704–11708. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15. Davis PJ, Smales CM, James DC. How can thermal processing modify the antigenicity of proteins? Allergy 2001;56:56–60. [DOI] [PubMed] [Google Scholar]