The construction and expression of phage display scFv library from the spleen cells of mice immunized with KGla cells

Objective:To construct a scFv library by phage display technique from the spleen cells of mice immunized with KGla cells. Methods: Three mice were immunized with KGla cells, and their spleen cells were harvested. The genes of VH and Vκ were amplified by RTPCR and a scFvphage display antibody library was constructed with the amplified V gene. The library was selected by using the KGla cells as panning antigen by 4 rounds of bindingelutionenrichment procedure, and the expression of the library were examined by SDSPAGE. Results: We were able to produce a scFv library containing 3×106individual clones which showed different patterns after digested with restriction endonuclease BstN I. It indicated that the capacity and diversity of the library is sufficient for screening specific scFv.The surface display expression of the library was also verified. Conclusion: A scFv library of antiKGla cell surface molecules has been constructed. It will be useful for isolating specific scFv against the cell surface molecules of hematopoietic progenitors.