An improved method for the purification of retinal S-antigen using selective hydrophobic adsorption chromatography.
This paper describes the use of phenyl-Sepharose CL-4B as a solid-phase hydrophobic adsorbent in the purification of S-antigen from protein extracts of bovine, porcine and human retina. Chromatographic conditions were ascertained whereby the majority of contaminating proteins were bound to the adsorbent leaving S-antigen in the liquid phase. In combination with size fractionation on Ultrogel AcA, the method conveniently yielded porcine and bovine S-antigen preparations up to 100% purity. Immunogenicity of purified S-antigens was verified by induction of experimental autoimmune uveoretinitis in albino Lewis rats. The method is preparative in scale, fast in performance and yields S-antigen in high purity and antigenic potency.
D000327:Adsorption; D000818:Animals; D000941:Antigens / Q000276:immunology / Q000302:isolation & purification*; D019393:Arrestin; D002417:Cattle; D002845:Chromatography; D005136:Eye Proteins / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D012685:Sepharose / Q000031:analogs & derivatives; D013552:Swine
D000941:Antigens; D019393:Arrestin; D005136:Eye Proteins; C020927:phenyl-sepharose; D012685:Sepharose