Affinity chromatography of mouse interferon: a modified purification procedure utilizing specifically purified antibodies.
Interferon preparations of a high degree of purity were obtained by a one-step procedure using affinity chromatography on specifically purified immunoadsorbent. The procedure consisted of binding interferon harvested from serum-free medium and purified by Zn-acetate precipitation and SP-Sephadex chromatography to CNBr-activated Sepharose 4B (Column No. 1). In the next step, antiinterferon globulin was purified by affinity chromatography on Column No. 1 with the bound interferon. In this way, antibodies against purified interferon, which were free from non-antibody components, were obtained. The purified antibodies were then coupled to CNBr-activated Sepharose 4B forming Column No. 2. The latter had a binding capacity of 125 800 mouse interferon units per 4.8 ml of gel. This capacity was not altered during an 8-month period of use. The gel was capable to bind interferons obtained from fibroblasts and leukocytes and, partially, from serum. The resulting purified, products were similar, i.e. they were not influenced by volume, interferon activity, or purity of the starting material. The electrophoretic profiles of the products had a similar shape irrespective of the origin of the starting material.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000302:isolation & purification; D001202:Ascitic Fluid / Q000166:cytology; D002846:Chromatography, Affinity / Q000379:methods*; D046508:Culture Techniques; D007372:Interferons / Q000096:biosynthesis / Q000097:blood / Q000302:isolation & purification*; D007739:L Cells / Q000378:metabolism; D007962:Leukocytes / Q000378:metabolism; D051379:Mice; D009522:Newcastle disease virus
D000888:Antibodies, Anti-Idiotypic; D007372:Interferons