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人红细胞凝集型单抗的制备与鉴定
Authors
吴婷
徐佩兰
+5 more
王生育
苏金华
邹明园
郑秀青
颜江华
Publication date
1 August 2018
Publisher
Doi
Cite
Abstract
目的制备人红细胞凝集型单克隆抗体并进行鉴定。方法以3种方式处理人血红细胞,获得红细胞免疫原,分别免疫Balb/c小鼠。应用杂交瘤技术,取免疫小鼠的脾细胞与Sp2/0骨髓瘤细胞融合,以凝集实验筛选分泌凝集型抗体的杂交瘤细胞,继而用有限稀释法克隆3次,并对杂交瘤细胞的稳定性进行测定。通过腹水法制备抗体,亲和层析法纯化抗体,对抗体的特性进行研究。结果 3种方式获得的免疫原均能建立有效的免疫反应,低渗加超声破碎法所获得的小鼠血清抗体效价最高。通过融合获得8株可稳定分泌抗体的杂交瘤细胞,其中,4A1分泌的抗体凝集效价最高。抗体可使4种人血型的红细胞凝集,无种属交叉凝集反应。结论有效制备了针对人红细胞的凝集性单抗,为其之后的应用研究奠定了基础。福建省省属公益类项目(2015R1036-3,2016R1034-2,2017R1036-3);;福建省科技计划(2014Y0082,2014Y0083
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Last time updated on 10/06/2020