[Generation of monoclonal antibody to GP73 based on antigen epitope].
OBJECTIVE
Preparation of monoclonal antibody (mAb) against GP73 protein.


METHODS
The N-terminal peptide (AAAERGAVELK) of GP73 protein was displayed on T7 phage, the recombinant phage was amplified and used as the immunogen to immunize mouse to produce antibody. The titer of the antiserum and the positive hybridoma clones which secreted the mAb against GP73 protein were detected by ELISA. The mAb specificity was assayed by ELISA and Western blot.


RESULTS
The high specificity mAb against GP73 protein was selected from the mouse immunized with the recombinant T7 phages displaying the epitope of GP73 by cell fusion and screening.


CONCLUSIONS
The appropriate protein epitope displayed on T7 phage could be used as alternative antigen to immunize animals to make specific antibody against the corresponding native protein.
Yao
Xi-ning
XN
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing 102206, China. xiningyao3579@yahoo.com.cn
Huo
Jing-rui
JR
Liu
Ying-fu
YF
Liu
Bao-cai
BC
Wu
Feng
F
Fan
Guo-cai
GC
Wang
Qing-ming
QM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005260:Female; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D000939:Epitopes; C410824:GOLM1 protein, human; D008565:Membrane Proteins