Glutamine and lysine as common residues from epitopes on α-lactalbumin and β-lactoglobulin from cow milk identified by phage display technology.

Cow milk is an important source of food protein for children; however, it could lead to allergy, especially for infants. α-Lactalbumin (α-LA) and β-lactoglobulin (β-LG) from whey protein make up a relatively high proportion of milk proteins and have received widespread attention as major allergens in milk. However, few studies have identified the epitopes of both proteins simultaneously. In this study, ImmunoCAP and indirect ELISA were first used for detection of sIgE to screen sera from allergic patients with high binding capacity for α-LA and β-LG. Subsequently, the mimotopes was biopanned by phage display technology and bioinformatics and 17 mimic peptide sequences were obtained. Aligned with the sequences of α-LA or β-LG, we identified one linear epitope on α-LA at AA 11–26 and 5 linear epitopes on β-LG at AA 9–29, AA 45–57, AA 77–80, AA 98–101, and AA 121–135, respectively. Meanwhile, the 8 conformational epitopes and their distributions of α-LA and β-LG were located using the Pepitope Server. Finally, glutamine and lysine were determined as common AA residues for the conformational epitopes both on α-LA and β-LG. Moreover, we found the addition of mouse anti-human IgE during the biopanning process did not significantly affect the identification of the epitopes. [Display omitted] [ABSTRACT FROM AUTHOR]