title: Interaction mechanism of mono-PEGylated proteins in electrostatic interaction chromatography.
abstract: 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-PEGylated, the binding site is not affected and the elution volume reduces due to the steric hindrance between PEGylated protein and ion-exchange ligand.
authors.affiliation: Bioprocess Engineering Laboratory, School of Engineering and Graduate School of Medicine, Yamaguchi University, Ube, Japan.
authors.forename: Mitsuyo
authors.initials: M
authors.lastname: Abe
authors.forename: Parvin
authors.initials: P
authors.lastname: Akbarzaderaleh
authors.forename: Masataka
authors.initials: M
authors.lastname: Hamachi
authors.forename: Noriko
authors.initials: N
authors.lastname: Yoshimoto
authors.forename: Shuichi
authors.initials: S
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002645:Chickens; D002852:Chromatography, Ion Exchange / Q000379:methods*; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism*; D055672:Static Electricity
chemical_list: D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; C421864:hen egg lysozyme; D009113:Muramidase