PEGylation of Proteins in Organic Solution: A Case Study for Interferon beta-1b
文献类型：期刊论文
| 作者 | Peng, Fei1,2; Wang, Yinjue1; Sun, Lijing1; Liu, Yongdong1; Hu, Tao1; Zhang, Guifeng1; Ma, Guanghui1; Su, Zhiguo1 |
| 刊名 | BIOCONJUGATE CHEMISTRY
 |
| 出版日期 | 2012-09-01 |
| 卷号 | 23期号:9页码:1812-1820 |
| 关键词 | multiple-sclerosis stability receptor lipase purification alpha/beta expression mechanism efficacy cloning |
| ISSN号 | 1043-1802 |
| 其他题名 | Bioconjugate Chem. |
| 中文摘要 | 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. |
| 英文摘要 | 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. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences |
| 类目[WOS] | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary ; Chemistry, Organic |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Chemistry |
| 关键词[WOS] | MULTIPLE-SCLEROSIS ; STABILITY ; RECEPTOR ; LIPASE ; PURIFICATION ; ALPHA/BETA ; EXPRESSION ; MECHANISM ; EFFICACY ; CLONING |
| 收录类别 | SCI |
| 原文出处 | |
| 语种 | 英语 |
| WOS记录号 | WOS:000308833600012 |
| 公开日期 | 2013-11-28 |
| 版本 | 出版稿 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/6424]  |
| 专题 | 过程工程研究所_研究所（批量导入） |
| 作者单位 | 1.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 |
| 推荐引用方式 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. |
入库方式： OAI收割
来源：过程工程研究所
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