Deng Ning, Su Kuan-Yuan, Wang Xun-Zhang, Long Qing-Xin, Yang Lin, Yu Zhou-Yao
State Key Laboratory for Biological Control, Biopharmaceutical Center in Zhongshan University, Guangzhou 510275, China.
Sheng Wu Gong Cheng Xue Bao. 2002 Sep;18(5):546-50.
Using of two-step integrating technology, transducted the H and L chain gene of humanized Fab fragment of anti-HB-sAg antibody into the genome of methylotropic yeast P. pastoris. Constructed a engineering yeast to produce humanized Fab fragment of the anti-HBsAg antibody. The Fab fragment was efficiently secreted into the medium at a concentration of 50-80 mg/L. The Fab fragment was purified from culturing supernatant of the recombinant yeas by affinity chromatography. The ELISA analysis showed the high affinity of the expressed humanized Fab fragment to the HBsAg.
采用两步整合技术,将抗HB - sAg抗体人源化Fab片段的重链和轻链基因导入甲基营养型酵母毕赤酵母基因组。构建了一株工程酵母以生产抗HBsAg抗体的人源化Fab片段。该Fab片段以50 - 80 mg/L的浓度高效分泌到培养基中。通过亲和层析从重组酵母的培养上清中纯化该Fab片段。ELISA分析表明所表达的人源化Fab片段对HBsAg具有高亲和力。