Purification and uveogenicity of retinal antigens
Fujino, Y.; Kawashima, H.; Okumura, A.; Mochizuki, M.
Nippon Ganka Gakkai Zasshi 91(4): 498-508
1987
ISSN/ISBN: 0029-0203 PMID: 3618399 Document Number: 298280
It has been suggested that some uveitogenic antigens were present in the retina. Recently, inter photoreceptor retinoid-binding protein (IRBP) was found to have uveitogenic activity similar to S-antigen (S-Ag). The present paper reports our purification method by which both IRBP and S-Ag were obtained in one process. Extract from bovine retinas was applied to gel chromatography (AcA 34 Ultrogel ) and fractions positive to anti-IRBP serum or anti-S-Ag serum were collected by double-diffusion-immuno-method. The IRBP-positive fractions were treated with ConA sepharose affinity chromatography and the S-Ag-positive fractions were treated with hydroxyapatite chromatography. Then after each fraction was tested by the double-diffusion-immuno-method using anti-IRBP serum or anti-S-Ag serum, only fractions with positive reactions were collected and used for IRBP or S-Ag, respectively. Two antigens obtained by this procedure were shown to have no cross reactivities by the double-diffusion-immuno-method. SDS-PAGE showed that molecular weights of IRBP and S-Ag were approximately 140K and 50K, respectively. Both IRBP and S-Ag caused experimental autoimmune uveoretinitis and pinealitis in Lewis rats, but IRBP was shown to be more potent than S-Ag by its earliler onset and higher frequency. (Acta Soc Ophthalmol Jpn 91:498-508, 1987).