Purification design and practice for pertactin, the third component of acellular pertussis vaccine, from Bordetella pertussis
文献类型：期刊论文
| 作者 | Li, Zenglan1,2; Zhang, Yan1; Wang, Qi1,2; Li, Zhengjun1,2; Liu, Yongdong1; Zhang, Songping1; Zhang, Guifeng1; Ma, Guanghui1; Luo, Jian1; Su, Zhiguo1,3 |
| 刊名 | VACCINE
 |
| 出版日期 | 2016-07-25 |
| 卷号 | 34期号:34页码:4032-4039 |
| 关键词 | Pertactin Acellular pertussis vaccine Surface electrostatic potential Purification Dual ion exchange chromatography |
| ISSN号 | 0264-410X |
| 英文摘要 | Development of acellular pertussis vaccine (aPV) requires purification of several components from Bordetella pertussis. While the components pertussis toxin (PT) and filamentous hemagglutinin (FHA) have been successfully purified, the third component, pertactin, proves to be a difficult target due to its very low concentration. In order to solve its purification problem, we performed the surface potential analysis with GRASP2 program. The results demonstrated that there are two major charge patches, one negative and one positive, which are located separately on this linear protein. For this special feature, we designed a dual ion exchange chromatography strategy including an anionic exchange and a cationic exchange process for separation of pertactin from the heat extract of B. pertussis. The initial anionic exchange chromatography concentrated the product from 1.7% to 14.6%, with recovery of 80%. The second cationic exchange chromatography increased the purity to 33%, with recovery of 83%. The final purification was accomplished by hydrophobic interaction chromatography, yielding a purity of 96%. The total recovery of the three columns was 61%. Characterization of the purified antigen was performed with CD, intrinsic fluorescence, HP-SEC and western-blot, showing that the purified protein kept its natural conformation and immune-reactivity. The rationally designed process proved to be feasible, and it is suitable for large-scale preparation of the third aPV component pertactin. (C) 2016 Elsevier Ltd. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Immunology ; Medicine, Research & Experimental |
| 研究领域[WOS] | Immunology ; Research & Experimental Medicine |
| 关键词[WOS] | OUTER-MEMBRANE PROTEIN ; ION-EXCHANGE SYSTEMS ; BINDING-AFFINITY ; IDENTIFICATION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000380604000018 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/21272]  |
| 专题 | 过程工程研究所_生化工程国家重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Jiangsu Natl Synerget Innovat Ctr Adv Mat, Nanjing 211800, Jiangsu, Peoples R China |
| 推荐引用方式 GB/T 7714 | Li, Zenglan,Zhang, Yan,Wang, Qi,et al. Purification design and practice for pertactin, the third component of acellular pertussis vaccine, from Bordetella pertussis[J]. VACCINE,2016,34(34):4032-4039. |
| APA | Li, Zenglan.,Zhang, Yan.,Wang, Qi.,Li, Zhengjun.,Liu, Yongdong.,...&Su, Zhiguo.(2016).Purification design and practice for pertactin, the third component of acellular pertussis vaccine, from Bordetella pertussis.VACCINE,34(34),4032-4039. |
| MLA | Li, Zenglan,et al."Purification design and practice for pertactin, the third component of acellular pertussis vaccine, from Bordetella pertussis".VACCINE 34.34(2016):4032-4039. |
入库方式： OAI收割
来源：过程工程研究所
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