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PEGylation of Proteins in Organic Solution: A Case Study for Interferon beta-1b
Alternative TitleBioconjugate Chem.
Peng, Fei1,2; Wang, Yinjue1; Sun, Lijing1; Liu, Yongdong1; Hu, Tao1; Zhang, Guifeng1; Ma, Guanghui1; Su, Zhiguo1
2012-09-01
Source PublicationBIOCONJUGATE CHEMISTRY
ISSN1043-1802
Volume23Issue:9Pages:1812-1820
AbstractConventional protein PEGylation is carried out in aqueous solution. However, some hydrophobic proteins seem to be stable in organic solution. In this study, a novel approach of PEGylating IFN-beta-1b in an organic solution of 2-butanol (2-BuOH) was investigated. Compared with protein PEGylation in aqueous solution, the overall modification yields increased more than 37%, while the yield of mono-PEGylated products could be increased by 36%. Furthermore, the PEGylated IFN-beta-1b, which was obtained in organic solution, demonstrated 18% more antiviral potency than those derived from aqueous solution. The PEGylation step could be directly connected to the previous protein separation step for process integration. Dynamic light scattering (DLS) and atomic force microscope (AFM) analysis revealed that IFN-beta-1b formed aggregates both in water and in 2-BuOH solutions. However, the aggregates were much smaller and more homogeneous in 2-BuOH than those in aqueous solution, thereby providing larger solvent accessible protein surfaces, which resulted in a more productive PEGylation process. In addition, the results of circular dichroism (CD), fluorescence spectra, and peptide mapping suggested that the increased bioactivity came from the difference in PEGylation site distribution due to solution environment that induced conformational discrepancy. The results of this study show that PEGylation of IFN-beta-1b in organic solution is a facile and efficient process, which might find applications for other hydrophobic proteins.; Conventional protein PEGylation is carried out in aqueous solution. However, some hydrophobic proteins seem to be stable in organic solution. In this study, a novel approach of PEGylating IFN-beta-1b in an organic solution of 2-butanol (2-BuOH) was investigated. Compared with protein PEGylation in aqueous solution, the overall modification yields increased more than 37%, while the yield of mono-PEGylated products could be increased by 36%. Furthermore, the PEGylated IFN-beta-1b, which was obtained in organic solution, demonstrated 18% more antiviral potency than those derived from aqueous solution. The PEGylation step could be directly connected to the previous protein separation step for process integration. Dynamic light scattering (DLS) and atomic force microscope (AFM) analysis revealed that IFN-beta-1b formed aggregates both in water and in 2-BuOH solutions. However, the aggregates were much smaller and more homogeneous in 2-BuOH than those in aqueous solution, thereby providing larger solvent accessible protein surfaces, which resulted in a more productive PEGylation process. In addition, the results of circular dichroism (CD), fluorescence spectra, and peptide mapping suggested that the increased bioactivity came from the difference in PEGylation site distribution due to solution environment that induced conformational discrepancy. The results of this study show that PEGylation of IFN-beta-1b in organic solution is a facile and efficient process, which might find applications for other hydrophobic proteins.
KeywordMultiple-sclerosis Stability Receptor Lipase Purification Alpha/beta Expression Mechanism Efficacy Cloning
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
DOI10.1021/bc300081f
URL查看原文
Indexed BySCI
Language英语
WOS KeywordMULTIPLE-SCLEROSIS ; STABILITY ; RECEPTOR ; LIPASE ; PURIFICATION ; ALPHA/BETA ; EXPRESSION ; MECHANISM ; EFFICACY ; CLONING
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary ; Chemistry, Organic
WOS IDWOS:000308833600012
Citation statistics
Cited Times:7[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/6424
Collection研究所(批量导入)
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
Peng, Fei,Wang, Yinjue,Sun, Lijing,et al. PEGylation of Proteins in Organic Solution: A Case Study for Interferon beta-1b[J]. BIOCONJUGATE CHEMISTRY,2012,23(9):1812-1820.
APA Peng, Fei.,Wang, Yinjue.,Sun, Lijing.,Liu, Yongdong.,Hu, Tao.,...&Su, Zhiguo.(2012).PEGylation of Proteins in Organic Solution: A Case Study for Interferon beta-1b.BIOCONJUGATE CHEMISTRY,23(9),1812-1820.
MLA Peng, Fei,et al."PEGylation of Proteins in Organic Solution: A Case Study for Interferon beta-1b".BIOCONJUGATE CHEMISTRY 23.9(2012):1812-1820.
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