Expression and activity identification of a human nasopharyngeal carcinoma I50 anti-idiotype antibody
10.3969/j.issn.1672-7347.2011.03.001
- VernacularTitle:全人源鼻咽癌抗独特型抗体I50的原核表达及活性鉴定
- Author:
Jiajia WANG
;
Yalin LI
;
Fengjie GUO
;
Guohua ZHOU
;
Guancheng LI
- Publication Type:Journal Article
- Keywords:
anti-idiotype antibody;
nasopharyngeal carcinoma;
prokaryotic expression;
Dot-ELISA
- From:
Journal of Central South University(Medical Sciences)
2011;36(3):185-191
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain I50 anti-idiotype antibody and identify its activity in vitro.Methods I50 anti-idiotype (Id) antibody gene was amplified from the template of fuse 5-I50 by PCR to construct a prokaryotic expression vector pET25b-I50. The expression of pET25b-I50 in E. coli BL21(DE3) was induced by isopropylthio-β-D-galactopyranoside (IPTG) and was confirmed by SDS-PAGE and Western blot with Ab1(FC2) monoclonal antibody and an anti-hexahistidine tag antibody. The method of dialysis refolding was used to restore the activity of I50 anti-Id antibody, which was measured by Dot-ELISA and lymphocyte proliferation assay. Results The recombinant vector was successfully constructed and the recombinant protein was successfully expressed and purified with 90% purity. The relative molecular weight of the expressed protein was 15 kD, which was in accordance with expectation. The activity of I50 anti-Id antibody could be restored and could promote the proliferation of lymphocyte in a dose-dependent manner. Conclusion These results suggested that I50 anti-Id protein vaccine is likely an option in the therapy against nasopharyngeal carcinoma in vivo.
