Abstract
A non-immunized human single chain variable fragment (scFv) library containing 2.5 × 107 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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This work was supported by the grant from program of the National Nature Science Foundation of China (Grant No. 30730068 and 31072122).
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Liu, Y., Chang, J., Chen, Y. et al. Construction of a human scFv antibody library with VH regions randomized and its application. Biotechnol Lett 34, 1203–1208 (2012). https://doi.org/10.1007/s10529-012-0898-3
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DOI: https://doi.org/10.1007/s10529-012-0898-3