title: Kinetic and stoichiometric analysis of the modification process for N-terminal PEGylation of staphylokinase.
abstract: Staphylokinase (SAK) is a therapeutic protein with promise for thrombolytic therapy of acute myocardial infarction. In this study, polyethylene glycol (PEG) aldehyde was used for N-terminal PEGylation of SAK to improve the pharmacological profiles of SAK. Due to the presence of the competitive PEGylation between the N terminus and the Lys residues, kinetic and stoichiometric analysis was carried out to investigate the process for the N-terminal PEGylation of SAK. To achieve this objective, size exclusion chromatography and tryptic peptide mapping were used to measure the PEGylation extent of SAK molecule and its specific amino acid residues, respectively.
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, People's Republic of China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Tao
authors.initials: T
authors.lastname: Hu
authors.forename: Yongdong
authors.initials: Y
authors.lastname: Liu
authors.forename: Guifeng
authors.initials: G
authors.lastname: Zhang
authors.forename: Guanghui
authors.initials: G
authors.lastname: Ma
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Su
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel / Q000379:methods*; D007700:Kinetics; D008666:Metalloendopeptidases / Q000737:chemistry*; D010449:Peptide Mapping / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D014357:Trypsin / Q000378:metabolism
chemical_list: D011092:Polyethylene Glycols; D014357:Trypsin; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus