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Primary structure and functional expression of heavy- and light-chain variable region genes of a monoclonal antibody specific for human fibrin. 1997
Z Song, and
Y Cai, and
D Song, and
J Xu, and
H Yuan, and
L Wang, and
X Zhu, and
H Lin, and
F Breitling, and
S Dübel
Institute of Hematology, Chinese Academy of Medical Sciences, Tianjin, China.
The immunoglobulin heavy- and light-chain variable region (VH and VK) genes were isolated from 8E5 hybridoma cells, which secreted monoclonal antibody against human fibrin by RT-PCR. An expression vector pOPE51-8E5 was constructed for the recombinant VH-VK scFv expression. The primary sequence of the variable regions was determined. Expression product was found in the periplasmic space and inclusion bodies by SDS-PAGE and immunoblotting. It was a 30 KDa single chain fragment (scFv) with the antigen-binding specificity of the parental monoclonal antibody. A light chain shuffling with an unspecific VL did not result in a loss of fibrin binding specificity.
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