%0 Journal Article
%T Production of Recombinant Humanized anti-HBsAg Fab Antibody by Fermentation<br>人源性抗HBsAg Fab抗体的发酵生产研究
%A DENG Ning
%A XIANG Jun Jian CHEN Wen Yin XIONG Sheng WANG Xun Zhang
%A SU Kuan Yuan
%A <br>邓宁
%A 向军俭
%A 陈文吟
%A 熊盛
%A 王珣章
%A 粟宽源
%J 生物工程学报
%D 2004
%I 
%X In order to produce recombinant human anti-HBsAg Fab antibody in Pichia pastoris, the recombinant yeast was fermented using fed-batch system in a 30 L bioreactor. The fermentation temperature was 30 degrees C, the pH was 5.0 approximately 5.3, and the DO was 20% approximately 30%. The recombinant Fab antibody was purified from crude culture supernatant by ion exchange and analyzed by SDS-PAGE and western blot and ELISA. When the absorbance (OD600) of broth reach 300 at the end of fed-batch phase, the induced phase was initiated. The results showed that recombinant human anti-HBsAg Fab antibody was high-level expressed in recombinant Pichia pastoris using a fed-batch fermentation system. Both chains of the Fab were successfully expressed upon methanol induction. After 192 h of induction, the expression level of recombinant Fab (soluble) reached 412 mg/L. The recombinant Fab antibody was purified effectively by ion-exchange chromatography from the fermentation supernatant to a purity of 95%. And the affinity activities of the purified recombinant Fab antibdy and fermentation supernatant were detected, and both of them showed high affinity activities. The results demonstrated that recombinant human anti-HBsAg Fab antibody could be high level produced by fed-batch fermentations in Pichia pastoris. Which can be efficiently used in industrial production.
%K HBsAg
%K recombinant Fab
%K fermentation
%K Pichia pastoris<br>乙型肝炎病毒
%K HBsAgFab
%K 抗体
%K 发酵
%K 毕赤酵母
%K 生物制品
%K 表面抗原
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=765B9542282E4248&yid=D0E58B75BFD8E51C&vid=A04140E723CB732E&iid=94C357A881DFC066&sid=FE6645F2371CA43C&eid=1F94F38CF0FA5258&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=3&reference_num=25