فهرست نشریات علمی دسترسی آزاد ایران

Background: The human heavy (VH) and light (VL) chain variable genes are amplified and randomly assembledtogether and cloned into the minor coat protein gene (g3p) of M13 bacteriophage. The resulting library of scFv isexpressed on the phage as g3p fusion protein. The high affinity specific scFv antibodies can be selected againsta key antigen using panning process. Our aim was development of scFv antibodies against P185 tumor antigenby recombinant phage antibody system and panning process.Methods: Antibody engineering technology was applied to lymphocyte mRNA of a non-immune donor and ascFv library was constructed. The library was panned against an immunodominant epitope of P185 which itsreactivity had been tested with sera from breast cancer patients. DNA fingerprinting of the scFvs selected thepredominated clones. These were then screened by ELISA.Results: A large library including high repertoires of VH and VL was constructed. DNA fingerprinting differentiateda number of clones. After selection against the immunodominant epitope of P185, nine clones were differentiatedincluding two predominated scFv antibodies. The predominated antibodies were bound to the correspondingepitope and produced a positive ELISA.Conclusion: The high affinity P185 specific scFv antibodies which were originated from human genes andbound specifically to the P185 epitope are valuable clinical agents and have the potential to be used in cancerimmunotherapy in which P185 overexpression and metastasis occurs.