Comparison of antigenicity and conformational changes to β-lactoglobulin following kestose glycation reaction with and without dynamic high-pressure microfluidization treatment.

Previous work indicated that conformational changes of β-lactoglobulin (β-LG) induced by dynamic high pressure microfluidization (DHPM) was related to the increase of antigenicity. In this study, β-LG glycated with 1-kestose and combined with DHPM decreased the antigenicity of β-LG. The antigenicity of control, β-LG-kestose (0.1 MPa) and β-LG-kestose (80 MPa) were 100, 79 and 42 μg/mL respectively. The molecular weight of β-LG conjugated to kestose increased from 18.4 to 19.6 kDa and its conformation scarcely changed. Conversely, combined with DHPM treatment (80 MPa), β-LG conjugated to kestose formed two conjugates with molecular weight of 18.8 and 19.8 kDa, respectively. Furthermore, the unfolding of β-LG as a result of the treatments is reflected by a decrease of intrinsic and synchronous fluorescence intensity and changes to the secondary structure. The conformational changes induced by DHPM and glycation treatments synergistically decrease the antigenicity of β-LG due to more masked or disrupted epitopes.