Guo-Qiang Bao, Xian-Li He
Department of General Surgery, Tangdu Hospital, Xi'an 710038, Shaanxi Province, China.
Methods Mol Biol. 2009;562:133-42. doi: 10.1007/978-1-60327-302-2_10.
We provide procedures for the panning of fully humanized Fab antibodies using guided selection. Human heavy and light chain genes are amplified. A parental light chain is cloned into a phage display vector and combined with the heavy chain library. After several rounds of panning, positive clones are identified and the heavy chain sequences that are recovered are combined with light chains for further selection by phage display. Human Fab antibodies are obtained that bind the same epitope as the parental antibody.
我们提供了使用导向选择淘选全人源化Fab抗体的方法。扩增人重链和轻链基因。将亲本轻链克隆到噬菌体展示载体中,并与重链文库组合。经过几轮淘选后,鉴定出阳性克隆,并将回收的重链序列与轻链组合,通过噬菌体展示进行进一步筛选。获得了与亲本抗体结合相同表位的人源Fab抗体。