RGNTF单抗与多抗的特性和抑制作用(简报)

继本实验室周明华等新近发现顶盖提取液(Te)中,有视网膜神经节细胞诱向因子(RGNTF)之后,为了进一步验证它的生物

CHARACTERISTICS AND INHIBITORY ACTION OF MONOCLONAL AND POLYCLONAL ANTIBODIES AGAINST THE RETINAL GANGLION NEURONOTROPHIC FACTOR (RGNTF)

We have immunized Balb/c mice and rabbits with a minute quantity of a 30 kD neuronotrophic factor which was isolated from the extract of newborn rat rectum (Te) by Phast System gel electrophoresis. Splenic cells from the immunized mice were hybridized with NS-1 mouse myeloma cells. Three clones were selected from 576 wells of hybridomas and were capable of secreting monoclonal antibodies specific to the retinal ganglion neuronotrophic factor (RGNTF-MAbs), namely A1, D3 and E8. Subtyping of the three monoclonal antibodies revealed that A1 and D 3 are IgG 3 and E 8 is IgM. They maintained secreting antibodies even after six months of culturing in vitro. In order to determine the specificities of these antibodies, we have used their ascites fluids containing antibodies at a different dilutions to study their effects on the survival of retinal ganglion cells in vitro. The results indicated that at the dilution ranges of 1:250 to 2000, all three monoclonal antibndies exhibited inhibition on the survival of retinal ganglion cells and the inhibition increased with increases in antibody concentrations; especially at a dilution of 1:250, the E 8 monoclonal antibody reaching 70% inhibition and A 1 and D 3 reaching 66% and 62% inhibition, respectively. Polyclonal antibodies from rabbits exhibited similar but weaker results of inhibition. We can conclude that the monoclonal and polyclonal antibodies can specifically inhibit the activity of the 30 kD retinal ganglion neuronotrophic factor.