Fermentation and Purification of HBscFv from P．pastoris on Pilot Scale

Journal of South China University of Technology (Natural Science Edition) ›› 2005, Vol. 33 ›› Issue (1): 79-83.

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Fermentation and Purification of HBscFv from P．pastoris on Pilot Scale

Xiong Sheng¹ Xie Qiu-ling² Wang Yi-fei¹ Deng Ning² Su Kuan-yuan³

1. Bio-medical Research and Development Center,Jinan Univ.,Guangzhou 5 10632,Guangdong,China;2.College of Bioscience and Biotechnology ,Jinan Univ.,Guangzhou 510632,Guangdong,China;3.Center of Infectious Diseases,the 458th Hospital of PLA,Guangzhou 510602,Guangdong,China

Received:2004-02-19 Online:2005-01-25 Published:2005-01-25
Contact: 熊盛(1972-)，男，博士，主要从事生物技术方面的研究 E-mail:xiongsheng99@263.net
About author:熊盛(1972-)，男，博士，主要从事生物技术方面的研究
Supported by:
国家自然科学基金资助项目(30371661，30400071)；广东省自然科学基金团队项目(粤科基办[2003]11)；广州市科技攻关项目(2003Z3-E0401)

Abstract

Abstract:

In order to produce soluble recombinant human anti-HBsAg single-chain Fv(HBscFv)in pilot scale,recombinant P.pastoris excreting HBscFv was inoculated in a 30-L fermentor to carry out a fed-batch culture.HB-scFv in the supernatant was then purified with a two-step procedure.namely,ion-exchange and immuno-affinity chromatography.It is found that,after the culture for 7 days,the cell density(OD6o0)of recombinant P．pastoris and the yield of target protein are respectively 334 and 260 mg/L．After the purification，HBscFv is collected with purity of 95%and a productivity of 171 mg/L.The results of bioassay demonstrate that the specific activity of HBscFv produced by P.pastoris is(2.52±0.17)μg^-2,without significant difference from that produced by E.coli..

Key words: HBsAg, single-chain Fv, P.pastoris, fermentation, purification

Xiong Sheng Xie Qiu-ling Wang Yi-fei Deng Ning Su Kuan-yuan. Fermentation and Purification of HBscFv from P．pastoris on Pilot Scale[J]. Journal of South China University of Technology (Natural Science Edition), 2005, 33(1): 79-83.

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Fermentation and Purification of HBscFv from P．pastoris on Pilot Scale

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