원문정보
초록
영어
We constructed a large human naive Fab library by phage display. Human B cells were isolated from PBL, bone marrow, lymph node, and spleen. The cDNAs encoding the light chain variable regions (VL) or heavy chain variable regions (VH) were synthesized by PCR from the B cells using the primers matching each of the functional germline alleles. The VL library was subcloned into the BstXI sites of pKB-Fab phagemid vector to generate a light chain library (1 X 108), and the VH library was subcloned into the SfiI sites of the light chain library to construct a Fab library (5 X 1010). For evaluation of the resulting Fab library, 200 VL (100 VK, 100 Vlamda) and 192 VH clones were randomly selected and analyzed for the sequences and expression in E.coli. Therapeutic antibodies were screened by panning against target antigens and characterized for their antigen binding affinities and biological activities.