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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 paper, human brain neuron-specific enolase (NSE) was isolated and purified by ion exchange and gel filtration chromatography. BALB / c mice were immunized with this antigen. Two hybridoma cell lines stably secreting monoclonal antibodies were obtained by the hybridoma technique and named as 581 and 3A8. Both hybridoma cells secreted IgG1 subclass antibodies. Indirect ELIsA experiments showed that 5B1 specifically reacted with NSE and did not cross-react with NSE isoenzyme NON. 3A8 had the same positive reaction with NSE and NNE. Western blotting results showed that 5B1 could specifically recognize NSE protein subunits with a molecular weight of 46 kD. The preparation of anti-NSE specific monoclonal antibody has application value for tumor diagnosis.