Rapid method for the preparation of an AGE-BSA standard calibrator using thermal glycation

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 37degreesC for 60 days. In the present study we attempted to reduce the duration of incubation to 4 days by increasing the temperature to 50degreesC. It is noteworthy that incubation at 50degreesC 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 37degreesC. 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. 11-15, 2005. (C) 2005 Wiley-Liss, Inc.