摘要
目的建立一种快速制备甲胎蛋白单克隆抗体(AFP-McAb)杂交瘤细胞系的方法。方法以常规和改良两种方式同时免疫8～12周Balb/c小鼠,取小鼠脾细胞与骨髓瘤细胞SP2/0融合并行次黄嘌呤、氨基蝶呤与胸苷选择性培养。用间接酶链免疫吸附试验法(ELISA)筛选阳性克隆株,并建立细胞系。结果在较短时间内,改良法能获得高效价的免疫抗体,与常规免疫法相比有极显著差异(P<0.01),同时改良法的细胞融合率和抗体阳性率也显著高于常规免疫法(P<0.001)。筛选获得的高效分泌抗人AFP-McAb的阳性杂交瘤细胞珠进行了初步鉴定。结论新的改良疗法优化了常规的McAb制备中动物免疫程序,并能成功筛选出AFP-McAb杂交瘤细胞系。
Objective： To establish a rapid method for preparing alpha-fetoprotein monoclonal antibody（AFP McAb）hybridoma cell line. Methods： Balb/c mice with 8-12 weeks were immunized by routine and modified methods respectively. The spleen cells from the immunized mice were fused with SP2/0 cell line of marrow tumor by technology of traditional hybridoma. The fused cells were selectively cultured with HAT medium. The positive cloning lines were screened by indirect enzyme-linked immunosorbent assay（ELISA）for establishing AFP- McAb cell lines. Results： The cell fusion rate and the McAb positive cell line rate in the modified method group were significantly higher than those in routine immune method group（P^0. 001）. The positive hybridoma cells secreting higher efficient McAb against human AFP were established. The preliminary identification for AFP- MeAb has been performed. Conclusions： The modified method is an optimized process of animal immunization method for preparing the AFP-McAb. The hybridoma cell line producing AFP-McAb has been established successfully.
出处
《生殖医学杂志》
CAS
2013年第5期360-363,共4页
Journal of Reproductive Medicine
基金
国家人口计生委科学技术研究所中央级公益性科研院所基本科研业务费专项(2010GJSSJKB01)
关键词
甲胎蛋白
杂交瘤
单克隆抗体
Alpha-fetoprotein
Hybridoma
Monoelonal antibody
