CAJ | 학술논문

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∶819 200.结论成功制备具有高效价、高特异性的抗pgp3多克隆抗体.
목적 표체병순화D형사안의원체질립단백pgp3,제비서원성다극륭항체.방법 유도표체pgp3단백,경곡광감태-S-전이매(GST)자주순화후면역BALB/c소서,제비다극륭항체,Western인적、세포면역형광감정항체적특이성,간접ELISA법측정항체효개.결과 득도순화적pgp3단백,획득서원성다극륭항체,경Western인적화세포면역형광감정항체가특이성결합pgp3단백,ELISA측정소득항체효개위1∶819 200.결론 성공제비구유고효개、고특이성적항pgp3다극륭항체.
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 enzyme-linked 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.