The dry thermal glycation method was used to conjugate D-glucose and D-fructose with bovine serum albumin. Reactions were conducted at 50-104oC for 30 min. Depending on temperature, different levels of substitution were achieved. In the case of D-glucose average substitution level of up to 53 mol glucose/mol BSA were obtained. D-Fructose turned out to be less reactive than D-glucose. The pH of the reaction mixture was also found to affect the efficiency of the glycation reaction. The levels of substitution were estimated using matrix assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF) and SDS-polyacrylamide gel electrophoresis. Glycated molecules, and a process of glycation itself are implicated in diabetes complications. Hypothetically bovine spongiform encephalopathy (BSE) was also derived from glycation.
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