NMDA受体NR2B亚单位特异性单克隆抗体的制备及人胚脑皮层NR2B蛋白表达的研究

Preparation of NMDA Receptor NR2B Subunit-specific Monoclonal Antibody and its Application in Detecting NR2B Protein in Cortex of Human Embryos

目的：制备NMDA受体NR2B亚单位特异性单克隆抗体，并对人胚脑皮层NR2B的表达作免疫印迹分析。方法：制备NR2BC末端934～1457氨基酸与GST的融合蛋白，并以此作为免疫原，经常规杂交瘤技术制备单克隆抗体。用该抗体对不同周龄的人胚脑皮层的NR2B蛋白作免疫印迹分析。结果：本研究制备的NR2B单克隆抗体能特异地识别180kDa的NR2B蛋白，并首次发现人胚脑皮层NR2B亚单位蛋白的表达。结论：成功制备了NR2B特异的单克隆抗体；12周龄的人胚脑皮层组织已有NR2B的表达，表达量随胎龄增大而增高。

Objective:To develop a monoclonal antibody (McAb) against NR2B subunit of NMDA receptor andto detect NR2B expression in cortex of human embryos by using the antibody. Methods: A fusion protein of GSTwith NR2B C-terminal (amino acids 934-1457) was prepared and used as an immunogen to make McAb byconventional hybridoma techniques. The cortex tissues of human embryos from different development stages wereimmunoblotted with the McAb. Results:The antibody specifically recognized a single band of approximate 180kDa of NR2B;and the expression of the NRZB subunit protein was found for the first time in human embryos.Conclusions: A McAb specifically against NRZB has been developed. The human embryo cortex expresses NR2Bat least as early as 12 weeks of pregnancy,the expression increases as embryos grow up.