摘要
中试规模生产可溶性重组人抗HBsAg单链抗体 (HBscFv) ,分泌表达HBscFv的巴氏毕赤酵母 (P .pastoris)工程菌在 30L发酵罐中进行补料分批培养 .上清中的HBscFv以离子交换及免疫亲和层析两步法进行纯化 .发现酵母工程菌经 7d补料分批培养后 ,6 0 0nm波长下的光密度值 (OD6 0 0 )达到 334,目的蛋白表达量为 2 6 0mg/L .纯化后 ,可获得纯度为95 %的HBscFv ,产量为 171mg/L .活性测定结果表明 ,HBscFv制品的比活性为 (2 5 2±0 17) μg- 1,与大肠杆菌来源的HBscFv无明显差异 .
In order to produce soluble recombinant human anti-HBsAg single-chain Fv (HBsc Fv) in pilot scale, recombinant P. pastoris excreting HBscFv was inoculated in a 30-L fermentor to carry out a fed-batch culture. HBscFv in the supernatan t 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 c ell density (OD 600) of recombinant P. pastoris and the yield of target protein are respectively 334 and 260 mg/L. After the purification, HBscFv is c ollected with a purity of 95% and a productivity of 171 mg/L. The results of bi oassay demonstrate that the specific activity of HBscFv produced by P. pastori s is (2 52±0 17) μg -1, without significant difference from that prod uced by E. coli..
出处
《华南理工大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2005年第1期79-83,共5页
Journal of South China University of Technology(Natural Science Edition)
基金
国家自然科学基金资助项目 (30 3716 6 1
30 4 0 0 0 71)
广东省自然科学基金团队项目 (粤科基办[2 0 0 3]11)
广州市科技攻关项目 (2 0 0 3Z3-E0 4 0 1)
