Native PAGE eliminates the problem of PEG-SDS interaction in SDS-PAGE and rovides an alternative to HPLC in 2 characterization of protein PEGylation
文献类型：期刊论文
| 作者 | Zheng, Chunyang Y.; Ma, Guanghui; Su, Zhiguo |
| 刊名 | ELECTROPHORESIS
 |
| 出版日期 | 2007-08-01 |
| 卷号 | 28期号:16页码:2801-2807 |
| 关键词 | native PAGE PEG-protein conjugates PEGylation SDS-PAGE |
| ISSN号 | 0173-0835 |
| 其他题名 | Electrophoresis |
| 中文摘要 | PEGylation of proteins has become an increasingly important technology in recent years. However, determination and characterization of the PEGylation products are problematic especially for the reaction mixture containing various modified proteins, unreacted PEG, and unmodified protein. A comparative study was carried out with two HPLC methods and two electrophoresis methods for characterization of the reaction mixture in PEGylation of HSA with PEG 5000, 10000, and 20000. RP-HPLC fails to give the correct information about the reaction of PEG 20000. Size-exclusion HPLC (SE-HPLC) produced very poor resolution on the PEG 5000 reaction. SDS-PAGE can run multiple samples of all PEGylation but the bands were smeared or broadened probably due to the interaction between PEG and SDS. On the other hand, native PAGE eliminates the problem of PEG-SDS interaction and provides better resolutions for all samples. Various PEGylated products and unmodified protein migrate differentially in native PAGE under nondenatured conditions. The results demonstrated that native PAGE could be a good alternative to HPLC and SDS-PAGE for the analysis of PEG-protein conjugates especially for characterization of the PEGylation mixture. |
| 英文摘要 | PEGylation of proteins has become an increasingly important technology in recent years. However, determination and characterization of the PEGylation products are problematic especially for the reaction mixture containing various modified proteins, unreacted PEG, and unmodified protein. A comparative study was carried out with two HPLC methods and two electrophoresis methods for characterization of the reaction mixture in PEGylation of HSA with PEG 5000, 10000, and 20000. RP-HPLC fails to give the correct information about the reaction of PEG 20000. Size-exclusion HPLC (SE-HPLC) produced very poor resolution on the PEG 5000 reaction. SDS-PAGE can run multiple samples of all PEGylation but the bands were smeared or broadened probably due to the interaction between PEG and SDS. On the other hand, native PAGE eliminates the problem of PEG-SDS interaction and provides better resolutions for all samples. Various PEGylated products and unmodified protein migrate differentially in native PAGE under nondenatured conditions. The results demonstrated that native PAGE could be a good alternative to HPLC and SDS-PAGE for the analysis of PEG-protein conjugates especially for characterization of the PEGylation mixture. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences |
| 类目[WOS] | Biochemical Research Methods ; Chemistry, Analytical |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Chemistry |
| 关键词[WOS] | SODIUM DODECYL-SULFATE ; POLYACRYLAMIDE-GEL-ELECTROPHORESIS ; MODIFIED SUPEROXIDE-DISMUTASE ; GLYCOL-MODIFIED PROTEINS ; CAPILLARY-ELECTROPHORESIS ; POLY(ETHYLENE GLYCOL) ; CHROMATOGRAPHY ; SEPARATION ; TERMINUS ; SITE |
| 收录类别 | SCI |
| 原文出处 | |
| 语种 | 英语 |
| WOS记录号 | WOS:000249286500002 |
| 公开日期 | 2013-10-15 |
| 版本 | 出版稿 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/3530]  |
| 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 100080, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zheng, Chunyang Y.,Ma, Guanghui,Su, Zhiguo. Native PAGE eliminates the problem of PEG-SDS interaction in SDS-PAGE and rovides an alternative to HPLC in 2 characterization of protein PEGylation[J]. ELECTROPHORESIS,2007,28(16):2801-2807. |
| APA | Zheng, Chunyang Y.,Ma, Guanghui,&Su, Zhiguo.(2007).Native PAGE eliminates the problem of PEG-SDS interaction in SDS-PAGE and rovides an alternative to HPLC in 2 characterization of protein PEGylation.ELECTROPHORESIS,28(16),2801-2807. |
| MLA | Zheng, Chunyang Y.,et al."Native PAGE eliminates the problem of PEG-SDS interaction in SDS-PAGE and rovides an alternative to HPLC in 2 characterization of protein PEGylation".ELECTROPHORESIS 28.16(2007):2801-2807. |
入库方式： OAI收割
来源：过程工程研究所
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