抗牛型结核分支杆菌单克隆抗体的制备和鉴定

运用杂交瘤细胞技术,以牛型结核分支杆菌免疫Balb/c小鼠,用其超声波裂解液筛选杂交瘤细胞,得到了2株能稳定分泌抗牛型结核杆菌抗体的杂交瘤细胞株,分别命名为5D4和4C3,并对它们的特性作了初步鉴定.经鉴定,2株单克隆抗体均为IgG1亚类.腹水经ELISA检测,效价可达1∶1×104~1∶1×105,同时分析了2株单抗的抗原位点,结果表明2株单抗针对不同的抗原位点.单抗的特异性试验结果显示,2株单抗与大肠杆菌,沙门氏菌,金黄色葡萄球菌等均没有反应,但都与人型结核分支杆菌有不同程度的交叉反应.并应用免疫胶体金技术制备牛结核杆菌胶体金快速诊断试纸条做应用性鉴定.该试纸条只与结核杆菌发生反应,而不与沙门氏菌、大肠杆菌、金黄色葡萄球菌、链球菌、李氏杆菌等发生交叉反应.该试纸条最低能检测出浓度为1×106个/mL的牛结核杆菌菌液,有较高的灵敏度.

Preparation and identification of monoclonal antibodies against Mycobaterium bovis

Two stable hybridoma cell lines(5D_4 and 4C_3) secreting monoclonal antibodies(McAbs) against Mycobaterium bovis were developed by fusion between the murine myeloma cell SP2/0 and spleen cells of BALB/C mice immunized with M.B.Their properties were identificated.The two cell lines belonged to class IgG_1.Indirect enzyme-linked immunosorbent assays(ELISA) were used to screen those hybridomas producing antibodies.The ELISA titers of McAbs were 1∶10~4 to 1∶10~5.Antigen dots and specific of the two McAbs were analysed.The two McAbs were against different antigen dots.The two McAbs were only positive against Mycobacterium tuberculosis,negative against Escherichia coli,Salmonella,Staphylococcus aureus.The two McAbs were applied to prepare the rapid detection papers of Mycobaterium bovis by colloidal gold immunochromatography.