摘要
视网膜神经节细胞诱向因子(RGNTF,Retinal ganglion neuronotrop-bic factor)是新发现的一神经生长因子,它能特异性地促进体外培养的视网膜神经节细胞的生长活性。作者用Phast System电泳实验证实,RGNTF的分子量为30kD蛋白质。并用之免疫动物,制备出3株抗RGNTF单克隆抗体,分别命名为A1、D3、E8。经鉴定杂交瘤细胞系染色体大多为端着丝点染色体,并有1～2条中部着丝点染色体。抗体的亚型鉴定A1、D3是IgG3型、E8是IgM型。用ELISA方法测定RGNTF-McAb与相应抗原结合的效价为10^(-4)～10^(-5)结果发现随抗体浓度的增强,抗体与抗原结合率也增加,以E8株结合率最高。本单抗与其它生长因子MGF、EGF无交叉反应,表明其相应抗原是一种新的生长因子。
Retinal ganglion neuronotrophic factor (RGNTF) is a newly discovered neural growth factor which can promote specifically the growth activity of retinal ganglion cells in vitro. We have shown by Phast System gel electrophoresis that RGNTF is a protein of molecular weight 30kD, which is usedias antigen to develop successfully 3 clones of hybridomas producing 3 monoclonal antibodies, namely Al, D3, and E8. Assessment of these clones of hybridomas reveals that the number of chromosomes in Al, D3, and E8 are 92, 89, and 98, respectively, and they are mainly teloc-chromosomes with 1-2 mefacentric chromosomes, Monoclonal antibodies from Al and D3 arc IgG3 subtype and F.8 is IgM subtype. The effecting binding value between these monoclonal antibodies and the antigen (at dilution from 10-4 to 10-5) reveals that the efficiency of binding between antibody and antigen increases with the increase of concentration of antibody with the E8 clone having the highest rate of binding. Our monoclonal antibodies do not crossrcact with both nerve, growth factor (NGF) and epidermal growth factor (EGF).These results suggest that RGNTF is a noval neural growth factor.
关键词
视网膜细胞
诱向因子
单克隆抗体
retinal ganglion cell
trophic factor
monoclonal antibody
assessment
growth factor.
