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Purification design and practice for pertactin, the third component of acellular pertussis vaccine, from Bordetella pertussis
Li, Zenglan1,2; Zhang, Yan1; Wang, Qi1,2; Li, Zhengjun1,2; Liu, Yongdong1; Zhang, Songping1; Zhang, Guifeng1; Ma, Guanghui1; Luo, Jian1; Su, Zhiguo1,3
2016-07-25
发表期刊VACCINE
ISSN0264-410X
卷号34期号:34页码:4032-4039
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.
关键词Pertactin Acellular Pertussis Vaccine Surface Electrostatic Potential Purification Dual Ion Exchange Chromatography
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine
DOI10.1016/j.vaccine.2016.06.029
收录类别SCI
语种英语
关键词[WOS]OUTER-MEMBRANE PROTEIN ; ION-EXCHANGE SYSTEMS ; BINDING-AFFINITY ; IDENTIFICATION
WOS研究方向Immunology ; Research & Experimental Medicine
WOS类目Immunology ; Medicine, Research & Experimental
项目资助者National Key Basic Research Program of China(2013CB733604) ; National High Technology Research and Development Program of China(2014AA021005 ; Natural Sciences Foundation of China(21306207 ; 2014AA021006) ; 21336010)
WOS记录号WOS:000380604000018
引用统计
文献类型期刊论文
条目标识符http://ir.ipe.ac.cn/handle/122111/21272
专题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
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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.
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