The Construction and Expression of Phage Display scFv Library from the Spleen Cells of Mice Immunized With B3HM Cells / 中国生物工程杂志

ABSTRACT

To construct a scFv library by phage display technique from the spleen cells of mice immunized with B3HM cells. Three mice were immunized with B3HM cells, and their spleen cells were harvested. The genes of VH and Vk were amplified by RT-PCR from the cDNA of the immunized spleen cells and a scFv-phage display antibody library was constructed. The capacity of library was measured,and the variety of the library was analyzed by digesting with restriction endonuclease BstNI.ScFv phage clones were randomly picked and identified phage-scFv clone by binding B3HM cells using immunofluorescein.A scFv library containing 5?106 individual clones which showed different patterns after digested with restriction endonuclease BstNI was produced. Individnal phage-scFv clone showed B3HM cells positive using immunofluorescein. A scFv library of anti-B3HM cell surface molecules has been constructed. It will be useful for finding out some novel genes of causing leukemia, and establishs the infarctate foundation of clarifying the pathogenesis of leukemiagenesis.