[Cloning and analysis of phage Fab antibodies of mouse male specific antigen].
To clone mouse phage antibodies against H-Y antigen from a phage antibody library, three cycles of affinity enrichment of the mouse phage antibody library with male spleen cells and two cycles of nonspecific absorption with female spleen cells were performed. The presence of mouse Fab on the phage surface was determined by ELISA and sequence analysis. 9 of 15 strains can bind to male spleen cells with the specific activity. Recombination rate of the phage antibody library clones is 60%. Sequence analysis of the PCR products of plasmid DNA of E5 clones show VH and Vkappa had common characteristics shared by other known variable region of antibodies. The mouse phage Fab antibody could be used for identifying H-Y antigen, and for the development of sex determination of early embryos in mammals.
Wang
Nai-Dong
ND
College of Veterinary Medicine, Hunan Agricultural University, Changsha, China.
Xue
Li-Qun
LQ
Xu
Dao-Jun
DJ
Yuan
An-Wen
AW
Deng
Zhi-Bang
ZB
Cui
Shu-Liang
SL
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006182:H-Y Antigen / Q000032:analysis; D007140:Immunoglobulin Fab Fragments / Q000235:genetics* / Q000276:immunology; D007518:Isoantibodies / Q000235:genetics* / Q000276:immunology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library
D006182:H-Y Antigen; C051256:H-Y antibody; D007140:Immunoglobulin Fab Fragments; D007518:Isoantibodies; D019151:Peptide Library