In this study, hen egg white lysozyme was used as a model protein for PEGylation using N-terminal specific mPEG propionaldehyde (PEG aldehyde) in presence of cyanoborohydrate. A method for the analytical-scale purification of PEG lysozyme was developed using an Agilent 1260 Infinity Bio-Inert LC System and an Agilent Poroshell 120 SB-C18 column. Analytical-scale Fraction Collector with peak-based fraction collection was employed to collect the purified PEG lysozyme. The fractions were then re-analyzed by RP HPLC and SEC to demonstrate the homogeneity of the purified PEG conjugate.
Analytical-scale purification of PEG lysozyme was developed using an Agilent Bio-LC and Poroshell column. PEGylated lysozymes were re-analyzed by SEC and RP HPLC. The results indicated that purified PEG lysozyme was homogenous.