Construction of a human scFv antibody library with VH regions randomized and its application |
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Authors: | Yunchao Liu Junbiao Chang Yumei Chen Bo Wan Yinbiao Wang Gaiping Zhang |
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Institution: | School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, People's Republic of China. |
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Abstract: | A non-immunized human single chain variable fragment (scFv) library containing 2.5 × 10(7) individual clones was constructed from antibody variable region genes of 200 non-immunized donors. ScFv gene repertories were generated by randomly combining rearranged variable regions of heavy chain (VH) and natural occurring light chain (VL) using overlapping extension PCR (OE-PCR). Five recombinant protein antigens from different species were successfully used to select specific binders. Phage ELISA showed that the recombinant phage particle could specifically bind to non-structural protein 1 of Avian influenza virus. This method can therefore efficiently generate a phage antibody library. |
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