Kinetic and thermodynamic analysis of ultra-high pressure and heat-induced denaturation of bovine serum albumin by surface plasmon resonance
Abstract
Purpose: To undertake comparative kinetic and thermodynamic analyses of the interaction of bovine serum albumin (BSA) with IgG pre-treated with ultra-high pressure (UHP) and moderate heat.
Methods: BSA solutions were processed at 100 – 600 MPa and 25 – 40 °C. We applied an optical biosensor based on surface plasmon resonance (SPR). The dissociation and association kinetics of antigen-antibody complexes were measured at different temperatures. By analyzing the resultant sensograms, the association rate constant (ka), dissociation rate constant (kd), equilibrium dissociation constant (KD), and thermodynamic parameters were calculated.
Results: The equilibrium disassociation constant, KD, ranged from a low value of 3.15 × 10−7 M (0.1 MPa, 25 °C) to a high value of 66.42 × 10−7 M (600 MPa, 55 °C). Increase in pressure and temperature led to decrease in the affinity of BSA for IgG. Pressure levels above 300 MPa promoted interactions between breakage of disulfide bonds, and the unfolding and aggregation of BSA.
Conclusions: These results show that the combination of UHP and moderate heat treatment cdecrease the allergenicity of BSA by changing their protein conformation.
Keywords: Ultra - high pressure, Bovine serum albumin, Surface plasmon resonance, Kinetics, Thermodynamics, Allergens
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