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

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 degrees 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 degrees C. It is noteworthy that incubation at 50 degrees 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 degrees 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.