沙眼衣原体质粒蛋白pgp3单克隆抗体制备及鉴定

发布时间：2018-03-27 06:23

本文选题：沙眼衣原体　切入点：分泌性蛋白　出处：《中国免疫学杂志》2015年05期

【摘要】：目的:原核表达沙眼衣原体(Ct)质粒分泌性蛋白pgp3,制备其单克隆抗体(mAbs)并鉴定其基本生物学特征。方法:构建pGEX-6p2-pgp3原核表达载体,在大肠杆菌中表达GST-pgp3融合蛋白作为抗原免疫BALB/c小鼠,取免疫后小鼠脾细胞与骨髓瘤细胞SP2/0融合,ELISA法筛选分泌抗pgp3蛋白mAbs的细胞株,对mAbs特异性、型别、类及亚类和效价进行鉴定。结果:GST-pgp3融合蛋白表达成功,且成功筛选出5株稳定分泌mAbs的杂交瘤细胞株,其中4株为分泌抗pgp3mAbs的杂交瘤细胞株(P1B3、P2A1、P2B6、P2C2),mAbs类型为IgG1/κ型,剩余1株为分泌抗GST蛋白的杂交瘤细胞株(P3B4),mAbs类型为IgG2b/κ型。交瘤细胞株P1B3、P2A1、P2B6、P2C2、P3B4培养上清中的mAbs效价分别为1∶6400、1∶3200、1∶12800、1∶6400和1∶6400。结论:成功制备出抗pgp3mAbs,为进一步研究Ct质粒蛋白pgp3功能及为Ct检测方法的建立奠定了基础。
[Abstract]:Objective: to express the secreted protein pgp3 of Chlamydia trachomatis (CT) plasmid, prepare its monoclonal antibody (mAbs) and identify its basic biological characteristics. Methods: the prokaryotic expression vector of pGEX-6p2-pgp3 was constructed. BALB/c mice were immunized with GST-pgp3 fusion protein expressed in Escherichia coli as antigen. The spleen cells and myeloma cell SP2/0 fusion Elisa were used to screen the cell lines secreting anti-#en3# protein mAbs, which were specific to mAbs and specific to the type of mAbs. Results the fusion protein was successfully expressed and five hybridoma cell lines secreting mAbs stably were successfully screened. Among them, 4 were hybridoma cell lines secreting anti-#en1#. The type of P2C2Abs was IgG1/ 魏 type. The remaining hybridoma cell line, P3B4, secreted anti-#en0# protein, was of IgG2b/ 魏 type. The mAbs titers in the supernatant of the supernatant of p1B3OP2B6, P2B6, P2C2P3B4 were 1: 4001: 32001128001w 6400 and 1: 64000.Conclusion: Anti-pgp3mAbs were successfully prepared for further study of the pgp3 function of Ct plasmid protein. And it lays a foundation for the establishment of Ct detection method.
【作者单位】：河北北方学院医学检验学院;邢台市人民医院检验科;
【基金】：校级青年基金资助项目(No.Q2013024)
【分类号】：R392

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