牙龈卟啉单胞菌特异性单克隆抗体的制备

目的 制备特异性识别牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)的单克隆抗体(monoclonal antibody,mAb)。方法 ①采用杂交瘤的方法制备m Ab,用P.g重组血凝素2(recombinant Hemagglutinin 2,r HA2)免疫BALB/c小鼠,间接ELISA筛选特异性识别P.g的杂交瘤细胞株;②间接ELISA检测mAb的效价及识别P.g的特异性、敏感性。结果 筛选到5株特异性识别P.g的m Abs,C10、F3、F3-2、G3和G3-2。其中,C10、G3和G3-2免疫球蛋白亚类为IgG2a型;F3和F3-2免疫球蛋白亚类为IgG1型。mAb C10、G3、F3和F3-2识别P.g的效价均1∶128 000;mAb G3-2识别P.g的效价1∶16 000;5株mAb识别r HA2的效价均1∶128 000。5株mAb均特异性识别5×105cfu/mL的P.g ATCC33277。结论 制备了5株特异性识别P.g的mAb,可用于各种免疫学技术,为开发P.g检测工具提供了条件,对牙周病的诊断和治疗有一定的意义。

Preparation of specific anti-Porphyromonas gingivalis monoclonal antibody

Objective To prepare specific anti-Porphyromonas gingivalis(P.g) monoclonal antibody(mAb).Methods (1)mAb was prepared through the hybridoma method,BALB/c mice were immunized with recombinant hemagglutinin 2(rHA2),and hybridoma cell lines were screened by indirect ELISA.(2)Indirect ELISA was used to detect the titer of mAb and to recognize the specificity and sensitivity of P.g.Results The five mAbs of C10,F3,F3-2,G3 and G3-2 were screened for specific recognition of P.g.Among them,C10,G3 and G3-2 immunoglobulin subclasses were the IgG2a type; F3 and F3-2 immunoglobulin subclasses were the IgG1 type.The titer of mAb C10,G3,F3 and F3-2 recognizing P.g was>1:128 000; the titer of mAb G3-2 recognizing P.g was>1:16 000; the titer of 5 mAbs recognizing rHA2 was>1:128 000.All specific mAbs recognize P.g ATCC33277 no less than 5×105 cfu/mL.Conclusion 5 mAbs for P.g specific recognition have been prepared,which can be used in various immunological techniques,providing conditions for the development of P.g detection tools.It is of a certain significance for the diagnosis and treatment of periodontal diseases.