CAJ | 학술논문

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pgp3 mAbs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pgp3 mAbs，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。
목적：원핵표체사안의원체（ Ct）질립분비성단백pgp3，제비기단극륭항체（ mAbs）병감정기기본생물학특정。방법：구건pGEX-6p2-pgp3원핵표체재체，재대장간균중표체GST-pgp3융합단백작위항원면역BALB／c소서，취면역후소서비세포여골수류세포SP2／0융합，ELISA법사선분비항pgp3단백mAbs적세포주，대mAbs특이성、형별、류급아류화효개진행감정。결과：GST-pgp3융합단백표체성공，차성공사선출5주은정분비mAbs적잡교류세포주，기중4주위분비항pgp3 mAbs적잡교류세포주（P1B3、P2A1、P2B6、P2C2），mAbs류형위IgG1／κ형，잉여1주위분비항GST 단백적잡교류세포주（P3B4），mAbs류형위IgG2b／κ형。교류세포주P1B3、P2A1、P2B6、P2C2、P3B4배양상청중적mAbs효개분별위1∶6400、1∶3200、1∶12800、1∶6400화1∶6400。결론：성공제비출항pgp3 mAbs，위진일보연구Ct질립단백pgp3공능급위Ct검측방법적건립전정료기출。
Objective:To express secreted protein-pgp3 of Chlamydia trachomatis(Ct)plasmid,produce monoclonal antibodies (mAbs)and identify their basic biological characteristics.Methods: Construction pGEX-6p2-pgp3 prokaryotic expression vector,then expressed GST-pgp3 fusion protein in E.coli as antigen used to immune BALB/c mice, spleen cells were fused with SP2/0 mouse myeloma cells.The hybridoma cell lines of screening mAbs were secreted by ELISA,and mAbs specificity,type,class and titer were identified.Results:GST-pgp3 fusion protein was successful expressed,5 strains stable hybridoma cell lines that secreted mAbs were screened out,including 4 strains secreted anti-pgp3 mAbs(P1B3,P2A1,P2B6,P2C2),mAbs type were IgG1/κ,the other strain secretion anti-GST mAbs(P3B4),mAbs type was IgG2b/κ.The titer of P1B3,P2A1,P2B6,P2C2,P3B4 were 1∶6 400,1∶3 200,1∶12 800,1∶6 400 and 1∶6 400 respectively.Conclusion:Successful prepared anti-pgp3 mAbs,and lay a foundation for further study the function of Ct plasmid protein pgp3 and the establishment of Ct detection method.