抗人IgG-抗HRP双特异性单克隆抗体细胞株的建立与鉴定

目的 制备人ＩｇＧ检测用双特异性单克隆抗体诊断试制。方法 用人ＩｇＧ免疫ＢＡＬＢ／ｃ小鼠脾细胞与抗ＨＲＰＭｃＡｂ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ＩｇＧ２ａ／ＩｇＧ１，余为ＩｇＧ１／ＩｇＧ１。培养上清与腹水效价分别为２＾７、１０＾０４以上，经ＨＲＰ亲和层析有２个蛋白吸ＢｓＭｃＡｂ存在于第２

The establishment and identification of cell line of anti-human IgG anti HRP bispecific monoclonal antibody

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