A new site-specific monoPEGylated β-lactoglobulin at the N-terminal: Effect of different molecular weights of mPEG on its conformation and antigenicity

Abstract A new method was investigated to decline the antigenicity of β-Lactoglobulin (β-LG) by site specifically conjugating β-LG at the N-terminus with 5 kDa and 10 kDa monomethoxy polyethylene glycol propyl aldehyde (mPEG-ALD). The optimal reaction conditions were molar ratio of 1:10 (β-LG:mPEG-ALD), reaction time for 16 h, and pH 5.0, and the content of mono-PEGylated β-LG was 51.3%. The results showed that mono-PEGylated β-LG with molecular mass of 23.2 kDa and 28.5 kDa. The peptide fragments of mPEG5kDa-ALD-β-LG produced the same sequence pattern of β-LG except for the absence of one peptides f(1–14), indicating that α-amino group at the N-terminal was selectively modified. Furthermore, the conformation of modified β-LG underwent into slight change. The antigenicity of mPEG5kDa-ALD-β-LG and mPEG10kDa-ALD-β-LG decreased from 144.4 μg/mL to 66.7 and 39.0 μg/mL respectively. It was speculated that the steric hindrance effect of PEG was the main reason for the decline of antigenicity of β-LG.