Prokaryotic expression and polyclonal antibody preparation of echovirus 18 capsid protein VP1
10.13200/j.cnki.cjb.004435
- VernacularTitle:埃可病毒18型衣壳蛋白VP1的原核表达及其多克隆抗体的制备
- Author:
LI Fei
- Publication Type:Journal Article
- Keywords:
Echovirus 18(E18);
Capsid protein VP1;
Prokaryotic expression;
Polyclonal antibodies
- From:
Chinese Journal of Biologicals
2025;38(03):297-301
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express capsid protein E18-VP1 of echovirus 18(E18) in prokaryotic cells and prepare the corresponding polyclonal antibodies,so as to lay a foundation for the immunological detection of E18 and the development of related kits.Methods The VP1 gene of E18-254 was cloned into vector pET30a to construct the recombinant plasmid pET30a-E18-254 VP1,which was transformed into competent cells of E.coli BL21(DE3) and induced by IPTG to obtain recombinant protein E18-VP1.The recombinant protein was then purified by nickel ion metal chelating chromatography.One female New Zealand white rabbit and five female BALB/c mice were immunized with the purified protein to prepare polyclonal antibodies.The titers of antibodies were determined by indirect ELISA,and the antibody specificity was detected by Western blot and indirect immunofluorescence assay(IFA).Results The purity of recombinant protein E18-VP1 was more than 95%after purification.The titers of rabbit and mouse anti-E18-VP1 polyclonal antibodies were 1:1 280 000 and 1:640 000,respectively,with high specificity.Conclusion The recombinant protein E18-VP1 was expressed in prokaryotic cells with high purity,and the polyclonal antibodies with high titer and specificity were obtained.
