Antigenic response of whey proteins and genetic variants of β-lactoglobulin — the effect of proteolysis and processing

Limited enzymatic hydrolysis trials of WPC-80 and β -lactoglobulin AA, AB and BB were performed using specific enzymes (trypsin, Neutrase, Corolase PP, Corolase PS) in a pH-stat under controlled conditions. The hydrolysates of WPC-80 and β -lactoglobulin were fractionated into high and low molecular fractions. Residual antibody binding activity of the peptides was dependent on the degree of hydrolysis (DH 2-20), but also on the enzyme used. Heat treatment affected the solubility and thereby the antigenic response. Dialysis influenced the antibody binding activity of the peptides. Tryptic hydrolysis of β -lactoglobulin AA was slower than for β -lactoglobulin BB and AB. Antigenic responses of the hydrolysates and fractions were measured by SLOT-BLOT and ELISA. SLOT-BLOT, a rapid screening method, was not able to differentiate the hydrolysates. The ELISA, being a more sensitive method, differentiated between the genetic variants, but was more time consuming. The lowest antigenicity was observed in the 1000–5000 Da fraction and β -lactoglobulin AA showed the lowest response.