Purification and characterization of human leukocyte interferon components.
Human leukocyte interferon, prepurified either by acid ethanol extraction or by affinity chromatography with antibodies, was further purified by gel filtration in the presence of sodium dodecyl sulfate. Interferon was eluted from gel filtration columns as an apparently homogeneous entity with a molecular weight of 26,600, resulting in an up to 50-fold additional purification during a single step. The antiviral activity could be further resolved into two components by hydroxylapatite adsorption chromatography. The isolated components (A and B) were distinguishable by isoelectric focusing and polyacrylamide gel electrophoresis. The apparent molecular weights were 20,000 to 16,000 and 16,000, respectively. No differences were detected in their susceptibility toward reduction of disulfide bonds by beta-mercaptoethanol. Both could be obtained on a preparative scale with minimal losses in biological activity.
D001665:Binding Sites; D002846:Chromatography, Affinity; D004220:Disulfides / Q000032:analysis; D006461:Hemolysis; D006801:Humans; D007372:Interferons / Q000302:isolation & purification / Q000378:metabolism; D007962:Leukocytes / Q000378:metabolism*; D008970:Molecular Weight; D010222:Parainfluenza Virus 1, Human; D011485:Protein Binding; D011487:Protein Conformation; D012967:Sodium Dodecyl Sulfate
D004220:Disulfides; D012967:Sodium Dodecyl Sulfate; D007372:Interferons