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本实验用Phast system电泳法,从大鼠视网膜神经节细胞诱向因子(retinal ganglion neuronotrophic factor,RGNTF)单克隆抗体(RGNTF-McAb)中,分离出一种免疫球蛋白lgG,免疫Balb/c小鼠,制备了抗RGNTF-McAb的独特型单克隆抗体。用MTT微量比色法,对该独特型单克隆抗体进行筛选。结果表明,独特型单克隆抗体G_4能特异性地和RGNTF单克隆抗体D_3结合,中和D_3对成视网膜细胞瘤(Rb)细胞的抑制作用;具有RGNTF的生物学效应,能促进Rb细胞的生长。用免疫组织化学方法证明,Rb细胞、大鼠胚胎视网膜节细胞、新生大鼠的视网膜节细胞都可对G_4独特型抗体发生阳性免疫反应,阳性颗粒较粗,主要分布在Rb细胞和视网膜节细胞的胞膜上,这些阳性颗粒可能是RGNTF的膜受体。所以我们认为,G_4单克隆抗体不仅是独特型抗体,而且是对位型的独特型抗体。它的产生对深入研究RGNTF的作用机制具有重要意义,亦有广阔的应用前景。
In this experiment, using Phast system electrophoresis, immunoglobulin lgG was isolated from rat retinal ganglion neuronotrophic factor (RGNTF) monoclonal antibody (RGNTF-McAb), immunized Balb / c small Mouse, a unique type of monoclonal antibody against RGNTF-McAb was prepared. This unique monoclonal antibody was screened by MTT microcolor assay. The results showed that the unique monoclonal antibody G_4 could specifically bind to RGNTF monoclonal antibody D_3 and neutralize the inhibitory effect of D_3 on retinoblastoma (Rb) cells. The biological effect of RGNTF can promote the growth of Rb cells . Immunohistochemistry showed that Rb cells, rat embryonic retinal ganglion cells and neonatal rat retinal ganglion cells were all positive for G_4-specific antibodies, and the positive particles were coarser and mainly distributed in Rb cells and retinal ganglion cells The positive particles may be RGNTF membrane receptors. So we think, G_4 monoclonal antibody is not only a unique type of antibody, but also the opposite type of unique antibodies. Its production is of great significance to further study the mechanism of action of RGNTF, and also has broad application prospects.