Purification, characterization, and attempts at isotopic labeling of mouse interferon.
Under optimal conditions which minimized the accumulation of extraneous proteins, interferon preparations were obtained in L cells containing from 2 x 10(4) to 5 x 10(4) units/mg of protein. The radiolabeled proteins were liberated simultaneously with interferon from cultures exposed to tritiated amino acids after viral stimulation and from corresponding controls, and were subsequently purified with the following results. Chromatography of interferon on carboxymethyl-Sephadex C-25 eliminated selectively unlabeled or poorly labeled proteins, resulting in a greater than sixfold increase in counts per minute per milligram of protein. Similarly purified control material harbored at least 12 times less tritium per milligram of protein than interferon, and the label was more diversely distributed among proteins of different molecular weights. On electrophoresis of interferon in polyacrylamide gels, labeled proteins were reduced further by a factor of at least 10 without loss in titer. Final purification was estimated at greater than 280-fold, representing a calculated specific activity of at least 1.4 x 10(7) units of interferon per milligram of protein.
D000179:Acrylates; D000650:Amnion; D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002586:Cesium; D002642:Chick Embryo; D002712:Chlorides; D002845:Chromatography; D002850:Chromatography, Gel; D003851:Deoxyribonucleases / Q000378:metabolism; D004586:Electrophoresis; D005782:Gels; D006801:Humans; D007372:Interferons / Q000096:biosynthesis / Q000302:isolation & purification; D007739:L Cells; D051379:Mice; D009522:Newcastle disease virus / Q000378:metabolism / Q000472:pathogenicity / Q000528:radiation effects; D010447:Peptide Hydrolases / Q000378:metabolism; D014176:Protein Biosynthesis; D012260:Ribonucleases / Q000378:metabolism; D013053:Spectrophotometry; D014316:Tritium; D014466:Ultraviolet Rays; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism / Q000472:pathogenicity / Q000528:radiation effects
D000179:Acrylates; D002712:Chlorides; D005782:Gels; D014316:Tritium; D002586:Cesium; D007372:Interferons; D003851:Deoxyribonucleases; D012260:Ribonucleases; D010447:Peptide Hydrolases