抗神经细丝蛋白单克隆抗体杂交瘤细胞株的建立与鉴定  

Establishment of monoclonal antibodies against neurofllament triplet proteins

作　　者：陈临[1] 邵文钊[1] 李希斌[1] 刘正梅[1] 郑玉书 历俊华[1] 赵景枫 

机构地区：[1]北京市神经外科研究所细胞生物室

出　　处：《中华神经外科杂志》1991年第3期208-210,I007,共4页Chinese Journal of Neurosurgery

摘　　要：本实验应用 Triton X-100处理,羟基磷灰石初步纯化和 DEAE 纤维素层析的方法,从牛脊髓中提取神经细丝蛋白,免疫小鼠,经细胞融合与鉴定,证明所能分泌抗神经细丝蛋白单克隆抗体杂交瘤细胞株特异性高,性质稳定,可以用于细胞学、组织学、病理学等方面的研究。Neurofilament(NF)triplet proteins were isolatedfrom bovine spinal cord by axonefloatation andisotoniclysis.The crude extraction were partiallypurified with hydroxyapatite gel by a batchwisemethod.Followed by DEAE-cellulose columnchromatography,the three components of NFproteins(NF-L,NF-M,NF-H)can be completelyseperated.The three bands of NF were immuno-stained on the transfered nitrocellulose paper.Two hybridoma clones secreting monoclonalantibodies against bovine neurofilament tripletproteins were prepared by fusion of spleen cellsfrom the mice immuninzed with crude andpurified bovine NF proteins with X63-Ag 8.653mouce myeloma cells,One hybridoma clone wasprepared by multi-well plate culture and anotherby soft agarose culture,They stained the neuronsand neuronal processces of human brain andspinal cord in frozen and paraffin embededsectinons.There two McAbs did not crossreactwith other intermidate filaments.The immunoglobulintypes of both McAbs were IgG2a(E_1)and IgM(D_2).

关 键 词：神经蛋白质类 单克隆抗体 细胞株 

分 类 号：R392.12[医药卫生—免疫学]