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A solid-phase adsorption method for PEGylation of human serum albumin and staphylokinase: preparation, purification and biochemical characterization
Suo, Xiaoyan1,2; Lu, Xiuling1; Hu, Tao1; Ma, Guanghui1; Su, Zhiguo1; Hu, T
2009-08-01
Source PublicationBIOTECHNOLOGY LETTERS
ISSN0141-5492
Volume31Issue:8Pages:1191-1196
AbstractA solid-phase adsorption method was developed to circumvent the disadvantage of the conventional liquid-phase PEGylation, i.e. the heterogeneity of the PEGylated products. The model proteins, human serum albumin (HSA) and staphylokinase (SAK), were adsorbed on the ion exchange chromatography media, followed by PEGylation with succinimidyl carbonate (SC)-mPEG5K and salt elution. Since PEGylation with SC-PEG5K alters the positive charge of the proteins, Q-Sepharose Big Beads and DEAE Sepharose Fast Flow were used for adsorption of HSA and SAK, respectively. Size exclusion chromatography and SDS-PAGE studies demonstrated that solid-phase PEGylation of proteins generate monoPEGylated proteins with the yield of 35-47%. Circular dichroism and intrinsic fluorescence studies showed that solid-phase PEGylation led to little conformational change of the proteins. Solid-phase PEGylation resulted in 35% loss in the biological activity of SAK, which is lower than the liquid-phase PEGylation (70%).
KeywordHuman Serum Albumin Pegylation Solid-phase Staphylokinase
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1007/s10529-009-9986-4
Indexed BySCI
Language英语
WOS KeywordHEMOGLOBIN
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000268002100010
Citation statistics
Cited Times:25[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/6609
Collection生化工程国家重点实验室
Corresponding AuthorHu, T
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Suo, Xiaoyan,Lu, Xiuling,Hu, Tao,et al. A solid-phase adsorption method for PEGylation of human serum albumin and staphylokinase: preparation, purification and biochemical characterization[J]. BIOTECHNOLOGY LETTERS,2009,31(8):1191-1196.
APA Suo, Xiaoyan,Lu, Xiuling,Hu, Tao,Ma, Guanghui,Su, Zhiguo,&Hu, T.(2009).A solid-phase adsorption method for PEGylation of human serum albumin and staphylokinase: preparation, purification and biochemical characterization.BIOTECHNOLOGY LETTERS,31(8),1191-1196.
MLA Suo, Xiaoyan,et al."A solid-phase adsorption method for PEGylation of human serum albumin and staphylokinase: preparation, purification and biochemical characterization".BIOTECHNOLOGY LETTERS 31.8(2009):1191-1196.
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