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目的:为了获得高效价和高特异性的抗Bf抗体。方法:以脆弱类杆菌的标准株免疫BALB/C小鼠,采用杂交瘤技术选择能稳定分泌抗脆弱类杆菌的杂交瘤细胞株。结果:建立了3株稳定的杂交瘤细胞株,ELISA法测定腹水McAb效价分别为1:6000,1:6000和1:12000,敏感性为102~103cfu,与其它杆菌、梭茵无交叉反应,竞争抑制试验表明3株McAb作用于细胞表面不同的抗原决定簇。将3株单克隆抗体两两混合制成单克隆抗体池用ELISA测定其敏感性较一株单克隆抗体提高2~4倍。结论:高效价和高特异性抗脆弱类杆菌的McAb对于研究脆弱类杆菌及其引起的感染性疾病有十分重要的意义。
Objective: To obtain high titer anti-Bf antibody with high specificity. Methods: BALB / C mice were immunized with the standard strain of Bacteroides fragilis, and the hybridoma cell lines that could stably secrete Bacteroides fragilis were selected by the hybridoma technique. Results: Three stable hybridoma cell lines were established. The titer of ascites McAbs were 1: 6000, 1: 6000 and 1: 12000, respectively. The sensitivity was 102-103 cfu. There was no cross reaction with other bacilli and serovar , Competitive inhibition test showed that three McAbs act on different antigenic determinants on the cell surface. Monoclonal antibody pools were prepared by mixing three monoclonal antibodies in two or two wells. The sensitivity of the monoclonal antibody pool was 2-4 times higher than that of a monoclonal antibody by ELISA. CONCLUSION: McAbs with high potency and high specificity against Bacteroides fragilis are of great significance for the study of Bacteroides fragilis and infectious diseases caused by it.