GFAP单克隆抗体制备及ELISA检测方法研究  

Preparation of Anti-GFAP Monoclonal Antibodies and Development of Double Antibody Sandwich ELISA for GFAP

作　　者：王珂[1] 张守涛[1] 郭亚楠[1] 

机构地区：[1]郑州大学生命科学学院,河南郑州450001

出　　处：《郑州大学学报（理学版）》2016年第2期95-100,共6页Journal of Zhengzhou University:Natural Science Edition

基　　金：国家自然科学基金青年科学基金资助项目(31501094);河南省基础与前沿技术研究项目(152300410032)

摘　　要：制备抗胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)单克隆抗体,并尝试建立双抗夹心ELISA检测方法,为出血性脑卒中早期诊断提供基础材料.用重组人GFAP作为抗原免疫BALB/C小鼠,通过杂交瘤技术制备抗GFAP单克隆抗体;用HRP标记单克隆抗体,ELISA和Western blot检测抗体的亚类、表位和特异性;采用ELISA技术制备GFAP检测试剂盒.获得了7株抗GFAP单克隆抗体,抗体亚类为Ig G2或Ig G1,抗体特异性好.抗体配对成功,ELISA检测重组人GFAP灵敏且稳定.Preparing monoclonal antibodies against GFAP and an ELISA kit for GFAP could provide reli- able tools for early diagnosis of hemorrhagic stroke. Hybridomacells were obtained by fusing myeloma cells （SP2/0） with spleen cells from BALB/C mice immunized with GFAP from human. After screened and subcloned, the seven hybridoma cell lines against human GFAP were obtained. The mcAbs from mice injected with hybridomacells were purified and conjugated to HRP. The subtype, epitopes and spe- cificity of monoelonal antibodies were determined by ELISA and Western blot. All hybridoma cells ob- tained were high specifically bound to GFAP, which were belonged to IgG1 or IgG2. Based on epitopes of GFAP mcAbs, the sandwich ELISA kit for GFAP was developed. All the results indicated that the high specific meAbs against GFAP had been prepared, and a rapid, sensitive and specific ELISA for recombi- nant GFAP detection was established successfully.

关 键 词：GFAP 出血性脑卒中 抗体 酶联免疫吸附检测 

分 类 号：Q813.2[生物学—生物工程] R392.12[医药卫生—免疫学]