摘要
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 。
ObjectiveIn order to prepare bispecific monoclonal antibody for detection of human IgGMethods8azaguanine(8AG) resistant hybridoma mutants keeping the ability of secreting anti HRPMcAb were used to fuse with spleen cells of BALB/c mice which had been immunized with human IgGResultsFive positive clones secreting antiIgGantiHRP BsMcAb were obtained The BsMcAb of one cell line was Ig2a/IgG1, the other were IgG1/IgG1 subclass The titer of the supernatant and ascitics of the five hybridhybridomas cell lines were over 2 -7 and 10 -5 respectively The BsMcAb were purfied by affinity chromatography The second peak can be measured BsMcAb The specifities of BsMcAb were analysed by ELISA and immunoblot, show that they can only react with human IgG The number of hybridhybridomas chromosomes is more than the parental ones After repeatedly frozen, thawed and passaged, the five hybridhybridomas were proved to be capable of secreting BsMcAb stablyConclusionThis method had higher sensitivity and simplicityIt can be used widely [
出处
《免疫学杂志》
CAS
CSCD
北大核心
2000年第3期225-227,238,共4页
Immunological Journal
