Construction of phage display antibody library of the special antigen responding to serum starved U251 cell
- VernacularTitle:U251细胞血清饥饿特异抗原的ScFv噬菌体抗体库构建
- Author:
Min YU
;
Hai SHANG
;
Deyong TAN
;
Guilin SUN
- Publication Type:Journal Article
- Keywords:
U251 cell;
Serum starvation;
ScFv;
Surface phage display;
Antibody library
- From:
Chinese Journal of Immunology
2000;0(08):-
- CountryChina
- Language:Chinese
-
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.
