[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 tectum (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 D3 are IgG3 and E8 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 antibodies 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 E8 monoclonal antibody reaching 70% inhibition and A1 and D3 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.
Zhou
M H
MH
Department of Anatomy, University of Hong Kong.
Ren
F
F
Zhao
L P
LP
Qiu
P X
PX
D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009414:Nerve Growth Factors / Q000276:immunology*; D009419:Nerve Tissue Proteins / Q000276:immunology*; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D012165:Retinal Ganglion Cells / Q000737:chemistry* / Q000166:cytology
D000911:Antibodies, Monoclonal; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins