Sequential Antibody Affinity Chromatography of Human Leukocyte Interferon

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Abstract
Antibodies to crude leukocyte inteferon (CIF) were bound to Sepharose 4B, and CIF was chromatographed. A relationship between recovered interferon and percentage binding of the antibodies to Sepharose 4B was observed. Highest recovery was obtained when only 50%-85% of the antibodies were bound. When CIF was subjected to normal affinity chromatography, specific activity of 1-5 .times. 106 interferon units (IFU) (69/19B-units)/mg protein was obtained. When this purified preparation was further chromatographed on a column mainly directed against normal cellular proteins, and then concentrated by normal affinity chromatography, a 10-fold increase in direct specific activity was obtained. The indirect specific activity is probably about 2 .times. 108 IFU/mg protein.