Construction of phage display antibody library of the special antigen responding to serum starved U251 cell / 中国免疫学杂志

ABSTRACT

Objective:To construct a phage display antibody library of special antigen responding to serum starved U251 cell,from which the serum responding gene and protein would be gotten.Methods:U251 cells were cultured in serum-absent midium for 48 h.Its protein was extracted and used to immunize BALB/c mice.Total RNA of the spleenocytes of immunized mice was extracted.VH and VL were amplified by RT-PCR and were linked into ScFv(Single chain fragment of variation) with a linker.ScFv was recombined to pCANTAB5E vector and was transformed to TG1 strain.Results:The library capacity was up to 3?106 cfu/L.A positive clone was identified from 8 random clones of this library.Conclusion:The special ScFv phage library is constructed successfully.It is the basis for screening special antibodies which can recognize serum responding protein.