CAS OpenIR  > 研究所(批量导入)
PEGylation of G-CSF in organic solvent markedly increase the efficacy and reactivity through protein unfolding, hydrolysis inhibition and solvent effect
Alternative TitleJ. Biotechnol.
Peng, Fei; Liu, Yongdong; Li, Xiunan; Sun, Lijing; Zhao, Dawei; Wang, Qingqing; Ma, Guanghui; Su, Zhiguo
2014-01-20
Source PublicationJOURNAL OF BIOTECHNOLOGY
ISSN0168-1656
Volume170Issue:1Pages:42-49
AbstractPrevious studies demonstrated that hydrophobic proteins could be PEGylated in organic phase rather than water phase. It is still not known what the difference is for a hydrophilic protein's PEGylation in these two different phases. In this study, granulocyte colony stimulating factor (G-CSF) was dissolved in neat dimethyl sulfoxide (DMSO) and was PEGylated. In comparison with the PEGylation in water solution, the PEGylation degree in the organic solvent increased by 33% and 42% for PEG-maleimide (MAL-PEG) and PEG-succinimidyl carbonate (SC-PEG) respectively. Structure analysis revealed that the protein was unfolded in DMSO, which could make the PEGylated sites of G-CSF easily accessible. The hydrolysis half-life in water solution was 40 min and 9 h for SC-PEG and MAL-PEG respectively. However, in DMSO solvent, PEGs were very stable and no hydrolysis could be detected. Stopped-flow demonstrated that the conjugation speed of G-CSF by MAL-PEG and SC-PEG in DMSO were 1.6 x 10(4) and 2 x 10(2) times faster than those in aqueous solution. The remarkable acceleration could mainly be attributed to an increase of protein nucleophilicity in DMSO. The results of this study could be referential to industrial application where the cost of PEG reagents and the speed of reaction on large scale are very important. (C) 2013 Elsevier B.V. All rights reserved.; Previous studies demonstrated that hydrophobic proteins could be PEGylated in organic phase rather than water phase. It is still not known what the difference is for a hydrophilic protein's PEGylation in these two different phases. In this study, granulocyte colony stimulating factor (G-CSF) was dissolved in neat dimethyl sulfoxide (DMSO) and was PEGylated. In comparison with the PEGylation in water solution, the PEGylation degree in the organic solvent increased by 33% and 42% for PEG-maleimide (MAL-PEG) and PEG-succinimidyl carbonate (SC-PEG) respectively. Structure analysis revealed that the protein was unfolded in DMSO, which could make the PEGylated sites of G-CSF easily accessible. The hydrolysis half-life in water solution was 40 min and 9 h for SC-PEG and MAL-PEG respectively. However, in DMSO solvent, PEGs were very stable and no hydrolysis could be detected. Stopped-flow demonstrated that the conjugation speed of G-CSF by MAL-PEG and SC-PEG in DMSO were 1.6 x 10(4) and 2 x 10(2) times faster than those in aqueous solution. The remarkable acceleration could mainly be attributed to an increase of protein nucleophilicity in DMSO. The results of this study could be referential to industrial application where the cost of PEG reagents and the speed of reaction on large scale are very important. (C) 2013 Elsevier B.V. All rights reserved.
KeywordPegylation Organic Solvent G-csf Dmso
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1016/j.jbiotec.2013.10.037
URL查看原文
Indexed BySCI
Language英语
WOS KeywordEGG-WHITE LYSOZYME ; DIMETHYL-SULFOXIDE ; DMSO ; DENATURATION ; NMR ; CONJUGATION ; CANDIDA ; PHENYL ; STATE ; CORE
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000329214700008
Citation statistics
Cited Times:6[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/8160
Collection研究所(批量导入)
AffiliationChinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
Recommended Citation
GB/T 7714
Peng, Fei,Liu, Yongdong,Li, Xiunan,et al. PEGylation of G-CSF in organic solvent markedly increase the efficacy and reactivity through protein unfolding, hydrolysis inhibition and solvent effect[J]. JOURNAL OF BIOTECHNOLOGY,2014,170(1):42-49.
APA Peng, Fei.,Liu, Yongdong.,Li, Xiunan.,Sun, Lijing.,Zhao, Dawei.,...&Su, Zhiguo.(2014).PEGylation of G-CSF in organic solvent markedly increase the efficacy and reactivity through protein unfolding, hydrolysis inhibition and solvent effect.JOURNAL OF BIOTECHNOLOGY,170(1),42-49.
MLA Peng, Fei,et al."PEGylation of G-CSF in organic solvent markedly increase the efficacy and reactivity through protein unfolding, hydrolysis inhibition and solvent effect".JOURNAL OF BIOTECHNOLOGY 170.1(2014):42-49.
Files in This Item:
File Name/Size DocType Version Access License
PEGylation of G-CSF (1599KB) 限制开放CC BY-NC-SAApplication Full Text
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Peng, Fei]'s Articles
[Liu, Yongdong]'s Articles
[Li, Xiunan]'s Articles
Baidu academic
Similar articles in Baidu academic
[Peng, Fei]'s Articles
[Liu, Yongdong]'s Articles
[Li, Xiunan]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Peng, Fei]'s Articles
[Liu, Yongdong]'s Articles
[Li, Xiunan]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.