Kinetic and stoichiometric analysis of the modification process for N-terminal PEGylation of staphylokinase |
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| Authors: | Wang Jun Hu Tao Liu Yongdong Zhang Guifeng Ma Guanghui Su Zhiguo |
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| Institution: | aNational Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, People’s Republic of China;bGraduate School, Chinese Academy of Sciences, Beijing 100190, People’s Republic of China |
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| 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. |
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