[Experiences with an isolation method for retinal S-antigen and interphotoreceptor retinoid-binding protein].
S-antigen (AgS) and interphotoreceptor retinoid-binding protein (IRBP) are two highly potent uveitopathogenic autoantigens of the retina frequently used to trigger autoimmune uveitis in animal trials.


OBJECTIVE
The aim of this study was the simultaneous isolation of both proteins from bovine retina, additionally avoiding time-consuming dialysis and employing prepacked, commercially available cartridges.


METHODS
Retina extract, obtained in the usual manner, was precipitated with ammonium sulfate. Both the desalting and the buffer exchange of dissolved precipitate were carried out using a Bio-Gel P6 column. Subsequent separation with Econo-Pac Q gave prepurified AgS and IRBP fractions. Further purification was realized predominantly with Econo-Pac Q, -HTP and -HIC (5-ml cartridges).


RESULTS
AgS and IRBP were isolated in high purity (sodium dodecylsulfate polyacrylamide gel electrophoresis, silver stain) from bovine retina using prepacked cartridges with yields of approximately 0.25 (for AgS) and 0.15 (for IRBP) mg/g retina, respectively.


CONCLUSIONS
The application of ready-to-use cartridges allows simultaneous isolation of AgS and IRBP in milligram amounts under simplified conditions. This approach might be of particular interest for small samples of retina.
Sych
F J
FJ
Klinik für Augenheilkunde, Friedrich-Schiller-Universität Jena.
Strobel
J
J
D000818:Animals; D019393:Arrestin / Q000302:isolation & purification*; D002417:Cattle; D005136:Eye Proteins / Q000302:isolation & purification*; D012160:Retina / Q000737:chemistry*; D012177:Retinol-Binding Proteins / Q000302:isolation & purification*; D014020:Tissue Extracts / Q000737:chemistry; D000099214:Retinol-Binding Proteins, Interstitial
D019393:Arrestin; D005136:Eye Proteins; D012177:Retinol-Binding Proteins; D014020:Tissue Extracts; D000099214:Retinol-Binding Proteins, Interstitial
Erfahrungen mit einem Präparationsverfahren für retinales S-Antigen und Interphotorezeptor-retinoid-bindendes Protein.