摘要
The combining site of OKT3 was modeled,and Human Ig LS1 and ND were selected asacceptors of CDRs of OKT3 VL and VH to construct a reshaping antibody against CD3.Bycomparing OKT3,LS1 and ND,with their own family sequences,some residues werechanged and the reshaping VL and VH genes were designed.The full VH gene was assem-bled in three steps with eight chemically synthesized oligonucleotide fragments using over-lapping PCR and sequenced.The VH gene was expressed as active protein and inclusion bod-ies in the vectors of pCOMB3 and pGEX-4T-1 by ELISA and Western blot analysis.
The combining site of OKT3 was modeled, and Human Ig LS1 and ND were selected as acceptors of CDRs of OKT3 VL and VH to construct a reshaping antibody against CD3. By comparing OKT3, LS1 and ND, with their own family sequences, some residues were changed and the reshaping VL and VH genes were designed. The full VH gene was assembled in three steps with eight chemically synthesized oligonucleotide fragments using overlapping PCR and sequenced. The VH gene was expressed as active protein and inclusion bodies in the vectors of pCOMB3 and pGEX-4T-1 by ELISA and Western blot analysis.
基金
Supported by National Natural Science Foundation of China
the High Technology Research and Development Programme of China.
