兔抗霍乱弧菌O139血清群rMSHA多克隆抗体的制备和鉴定

Preparation and identification of polyclonal antibodies against rMSHA from Vibrio cholerae O139 serogroup

目的制备兔抗霍乱弧菌O139血清群rMSHA多克隆抗体。方法用经Ni-NTA亲和层析法纯化的重组蛋白(rMSHA)免疫日本大耳兔,收集免疫兔血清。Western Blot检测重组蛋白的免疫原性;ELISA测定该抗体的效价。结果兔抗rMSHA抗体能与重组蛋白发生特异性反应;间接ELISA法测定该抗体效价为1∶25 600。结论成功地制备了兔抗rM-SHA抗体,该抗体能够识别原核表达的重组蛋白rMSHA,为研制霍乱弧菌疫苗及相关诊断试剂盒奠定了基础。

To prepare and identify the polyclonal antibodies against rMSHA from Vibrio Cholerae O139 serogroup, the fusion protein rMSHA purified by Ni-NTA affinity chromatograthy with adjuvants were used to immunize rabbit to obtain polyclonal antibodies. The immunogenicity of the recombinant protein was detected by Western Blot. And ELISA was applied to demonstrate the titer of antiserum in the immunized rabbit. It was shown that the specific rabbit antibody against the purified rMSHA fusion protein could combine with rMSHA fusion protein conformed by Western blotting, and the titer of antiserum demonstrated by ELISA was 1 ： 25 600. It is evident that fusion protein rMSHA with satisfactory immunoreactivity and antigenicity can be used as an antigen in cholerae genetic engineering vaccine and laboratory detection kit.