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目的制备人 Ig G检测用双特异性单克隆抗体诊断试剂。方法用人 Ig G免疫 BAL B/c小鼠脾细胞与抗HRPMc Ab杂交瘤细胞 HAT敏感株进行第 2次融合。结果获得 5株能稳定分泌抗人 Ig-抗 HRP的双特异性单克隆抗体的杂交-杂交瘤细胞 ,分泌的抗体亚类其中 1株为 Ig G2 a/Ig G1,余为 Ig G1 /Ig G1 。培养上清与腹水效价分别为 2 - 7,10 - 5以上 ,经 HRP亲和层析有 2个蛋白吸收峰 ,Bs Mc Ab存在于第 2峰。用 EL ISA法及免疫印迹试验证明 :Bs Mc Ab只与人 Ig G有特异性反应。杂交 -杂交瘤细胞连续 3月培养和反复冻存 ,复苏后仍能稳定保持分泌 Bs Mc Ab的能力。结论该方法制备简单 ,灵敏高度 ,具较好的应用前景
Objective To prepare bispecific monoclonal antibody diagnostic reagents for human Ig G detection. Methods BALB / c mice immunized with human Ig G were spleen cells fused with HAT-sensitive strain of HRPMc Ab hybridoma cells for the second time. Results Five hybridoma hybridomas secreting anti-human Ig-anti-HRP bispecific monoclonal antibodies were obtained. One of them was Ig G2 a / Ig G1 and the rest was Ig G1 / Ig G1 . The culture supernatants and ascites titers were above 2 - 7 and 10 - 5, respectively. There were 2 protein absorption peaks by HRP affinity chromatography, and Bs Mc Ab was present at the 2nd peak. EL ISA method and western blotting showed that Bs Mc Ab only reacted specifically with human Ig G. Hybridoma-hybridoma cells were cultured and frozen repeatedly for 3 months. After resuscitation, the ability of secreting Bs-McAbs remained stable. Conclusion The method is simple and sensitive to prepare with good application prospects