Title: Expression of recombinant PEB1 protein of campylobacter jejuni and preparation of monoclonal antibodies against it
Abstract: Objective To express recombinant PEB1 protein of campylobacter jejuni(CJ)and prepare,identify monoclonal antibodies against it.Methods E.coli BL21(DE3) was raised and induced to express the recombinant CJ PEB1 protein.The BALB/C mice were routinely immunized by the immunogen PEB1 protein emulsified with identical volume adjuvant,then the monoclonal antibodies against PEB1 protein were prepared.The splenocytes tissue were gathered and fused with SP2/0 cells,then the hybridoma cell lines were established.The hybridoma cells were used to induce ascites by intraperitoneal injection in BALB/C mice,then the antibody titer was detected with indirect ELISA and Double agar diffusion test,and the antibody specificity was examined by Western-blot and Slide agglutination test.Results The recombinant PEB1 protein with high purity was obtained,and the relative molecular weight of which was identical with theory value.Two hybridoma cell lines which secreted monoclonal antibodies steadily were obtained.Isotypes of these two cell strains showed that they belong to IgG1 and IgG3 respectively.The indirect ELISA showed that the antibodies titer of ascites induced by the two hybridoma cell lines were respectively 8×104 and 5×104,and the Double agar diffusion showed it was 1∶16.The monoclonal antibodies could specifically recognized PEB1 protein demonstrated by Western-blot.Slide agglutination test showed the ascites induced by two hybridoma cell lines appeared agglutinative mixing with CJ,but not with E coli,Bacillus dysenteriae,Bacillus typhosus and et al.Conclusion Two hybridoma cell lines stably secreting monoclonal antibodies against CJ PEB1 protein can be established,and the monoclonal antibodies against CJ PEB1protein can be prepared successfully,which provids a basis for the detection of CJ.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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