摘要
本文采用离子交换和凝胶过滤层析技术，分离纯化了人脑神经元特异性烯醇化酶（ＮＳＥ）．并以此为抗原免疫ＢＡＬＢ／ｃ小鼠。通过杂交瘤技术，获得了两株稳定分泌单克隆抗体的杂交瘤细胞株，命名为５８１和３Ａ８。两株杂交瘤细胞均分泌ＩｇＧ１型亚类抗体。间接ＥＬＩｓＡ实验显示：５Ｂ１与ＮＳＥ有特异性反应，与ＮＳＥ的同工酶──非神经元待异性烯醇化酶（ＮＮＥ）不发生交叉反应；３Ａ８与ＮＳＥ和ＮＮＥ有同等强度的阳性反应。免疫印迹实验结果表明：５Ｂ１能够特异性地识别分子量为４６ＫＤ的ＮＳＥ蛋白亚基。抗ＮＳＥ特异性单克隆抗体的制备，对于肿瘤诊断有应用价值。
In this study,Neuron-specific enolase(NSE)was purified from the human brain
tissue throuhg ion-exchange and gel fiItration chromatography.The prepared NSE was used as
antigen for immunization of Balb/c mice. Hybridoma technique was employed to raise
monoclonal antibodies against NSE. Two cell lines capable of steady produc-tion of monoclonal
antibodies were established and they were designated 5B1 and 3A8 respectively.Monoclonal
antibodiessecreted by both of the hybridoma cell lines belong to IgG 1 subelass.Indirect ELISA
results showed that 5B1 reacted onlywith NSE and had not cross-reacting with NNE.3A8 reacted
with NSE as well as NNE. Immunoblotting results indicatedthat 5B1 recognized NSE subunit
with a molecular weight of 46KD. The preparation of specific monoclonal antibodies forNSE have
potential utility in the diagnosis of tumors.
出处
《免疫学杂志》
CAS
CSCD
北大核心
1994年第3期191-193,共3页
Immunological Journal
