Purification of the major species of human leucocyte interferon with the help of a monoclonal antibody.
A mouse hybridoma cell line has been isolated, which secretes a monoclonal antibody (HBA) specific of human leucocyte (alpha) interferon. This monoclonal antibody neutralizes biological activities (cellular and antiviral) of most of the molecular species of alpha interferon produced by human leucocytes. Bound to Sepharose, it has been used as an immunoadsorbent (HBA-Sepharose) to purify human leucocyte interferon. Human leucocyte interferon can be purified to homogeneity by a two-step chromatography on the immunoadsorbent. A high recovery (70-90%) of purified leucocyte interferon is obtained from crude or partially purified preparations, with a specific activity of 2,8 X 10(8) IU/mg of protein. Analysis of the purified proteins by SDS-gel electrophoresis and silver staining shows that the purified human leucocyte interferon consists of three species of molecular weight: 18 000-21 000 and 27 000. This simple and rapid technique can be applied for the preparation of homogeneous leucocyte interferon on a large scale.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007163:Immunosorbent Techniques; D007370:Interferon Type I / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008970:Molecular Weight
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007370:Interferon Type I