Radiation-Induced Modifications in Bovine Serum Albumin in Saline Solutions Under E-Beam Irradiation

Electron beam irradiation, extensively used for suppressing a wide range of pathogens contaminating food products, pharmaceuticals and biological raw materials, inevitably damages the surrounding proteins, stripping the product of its essential nutritional and functional properties. This issue can be addressed by adjusting the electron beam irradiation dose, bearing in mind the concentration of proteins in the product since it can affect the rate of radiation-induced modifications in proteins. The study investigates the impact of 7.5 MeV electron-beam irradiation on modifications in bovine serum albumin (BSA) molecules in 0.9% NaCl solution in the concentration range of 0.5–70 mg/mL, encompassing a wide range of protein concentrations in food products, pharmaceuticals and biological raw materials. Conformational changes and aggregation of BSA were evaluated using UV–Vis spectrophotometry at λ = 350 nm. Peptide bond rupture in protein native structures was assessed by performing HPLC-MS/MS analysis after trypsin hydrolysis using three selected peptides located in different domains of the BSA amino acid sequence. It was found that the rate of radiation-induced modifications increased with an increase in the irradiation dose but decreased markedly as BSA concentration increased. While at the BSA concentration of 0.5 mg/mL over 87% of BSA molecules underwent peptide bond rupture under irradiation with a dose of 5 kGy, a two-fold increase in the BSA concentration and irradiation dose enabled bond rupture in only 20% of BSA molecules. Our experimental approach resulting in the development of the dose and concentration model allows us to quantify the degree of radiation-induced protein modifications depending on the irradiation dose and protein concentration in food products, pharmaceuticals and biological raw materials.