沙眼衣原体质粒蛋白pgp3多克隆抗体的制备及鉴定

Preparation and identification of polyclonal antibody against plasmid-encoded pgp3 protein of Chlamydia trachomatis

目的表达并纯化D型沙眼衣原体质粒蛋白pgp3，制备鼠源性多克隆抗体。方法诱导表达pgp3蛋白，经谷胱甘肽-S-转移酶（GST）磁珠纯化后免疫BALB／c小鼠，制备多克隆抗体，Western印迹、细胞免疫荧光鉴定抗体的特异性，间接ELISA法测定抗体效价。结果得到纯化的pgp3蛋白，获得鼠源性多克隆抗体，经Western印迹和细胞免疫荧光鉴定抗体可特异性结合pgp3蛋白，ELISA测定所得抗体效价为1：819200。结论成功制备具有高效价、高特异性的抗pgp3多克隆抗体。

Objective To express and purify pgp3 protein of Chlamydia trachomatis serovar D and prepare mouse anti-pgp3 polyclonal antibody. Methods The pgp3 protein was induced to be expressed and purified by glutathione S-transferase （GST） Magbeads. Then the purified pgp3 was injected into BALB/c mice to prepare polyclonal antibody. Western blot and immunofluorescence were used to identify the specificity of the antibody. The titer of the antibody was tested by indirect enzymeqinked immunosorbent assay （ELISA）. Results The pgp3 protein was successfully expressed and purified. The mouse anti-pgp3 polyclonal antibody was prepared. Results of Western blot and immunofluorescence indicated that it was able to specifically bind to the pgp3 protein. The titer of the polyclonal antibody was 1 .＂ 819 200 which was tested by ELISA. Conclusion The anti-pgp3 polyclonal antibody with high specificity and high titer is successfully obtained, which could be useful to the researches on pathogenicity of Chlamydia trachomatis.