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Interaction mechanism of mono‐PEGylated proteins in electrostatic interaction chromatography
Author(s) -
Abe Mitsuyo,
Akbarzaderaleh Parvin,
Hamachi Masataka,
Yoshimoto Noriko,
Yamamoto Shuichi
Publication year - 2010
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201000013
Subject(s) - pegylation , chemistry , lysozyme , chromatography , polyethylene glycol , peg ratio , steric effects , ion chromatography , elution , bovine serum albumin , affinity chromatography , ligand (biochemistry) , covalent bond , biochemistry , enzyme , stereochemistry , organic chemistry , receptor , finance , economics
The retention and binding mechanisms in electrostatic interaction‐based chromatography (ion‐exchange chromatography) of PEGylated proteins (covalent attachment of polyethylene glycol chains to protein) were investigated using our previously developed model. Lysozyme and bovine serum albumin were chosen as model proteins. The retention volume of PEGylated proteins shifted to lower elution volumes with increasing PEG molecular weight compared with the non‐modified (native) protein retention volume. However, PEGylation did not affect the number of binding sites appreciably. The enzyme activity of PEGylated lysozyme measured with a standard insoluble substrate in suspension decreased considerably, whereas the activity with a soluble small‐molecule substrate did not drop significantly. These findings indicate that when a protein is mono‐PEG‐ylated, the binding site is not affected and the elution volume reduces due to the steric hindrance between PEGylated protein and ion‐exchange ligand.