Proteomic and immunochemical approaches to understanding the glycation behaviour of the casein and β-lactoglobulin fractions of flavoured drinks under UHT processing conditions.

Dairy technology used to produce sweetened milk products might introduce additional advanced glycation end products (AGEs) into the diet. These molecular messengers are linked to detrimental health effects. Using a model accurate to the thermal treatment, reducing sugars, main protein content, and prolonged storage of ultra-high-temperature-sterilized (UHT) milk, we studied the behaviour of milk proteins during glycation. Two-dimensional electrophoresis (2-DE) profiles and western blots of glycated total casein revealed the major contributions of α-casein and β-casein and the relatively minor contributions of κ-casein towards the formation of N-carboxymethyllysine (CML)-positive aggregates. Glycated κ-casein had the lowest furosine (FUR), 5-hydroxymethylfurfural (HMF) and AGEs content. Conversely, the α-casein fraction demonstrated a high susceptibility to glycation, having the highest FUR, HMF and AGE levels. The gel-filtration elution profiles and the corresponding fraction fluorescence revealed that glycated casein aggregates were highly fluorescent, while the β-lactoglobulin glycation profile was similar to that of bovine serum albumin, and fluorescence was detected mainly in tetramers. Although CML is not a cross-linking AGE, it was only detected in large molecular aggregates and not in glycated monomers. Our results also indicate that in casein, glycation-induced changes in the UHT conditions were less deleterious than the subsequent 90 day storage period.