Radioactive human lymphoblastoid interferon. One-step purification, regulation of heterogeneous species production and its use for radioimmunoassay.
Human lymphoblastoid interferon (alpha type), labeled with [3H]leucine added to virus-induced Namalwa cells, was purified quantitatively and in one step from the culture fluid by immune precipitation. The material showed, upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, only four radioactive bands with molecular weights ranging from 17000 to 21000, which coincided well with interferon activity. They coincided also with the four interferon protein bands in the electropherogram of unlabeled interferon purified by a different method. The purity of the labeled interferon was ascertained also by polyacrylamide gel electrophoresis in the absence of dodecyl sulfate. Pulse-labeling of interferon with [3H]leucine for 1 h at various times after induction indicated that the cells always synthesized and secreted the four interferon species in parallel during the interferon production period. Competitive radioimmunoassay for human interferon alpha was achieved by the use of purified radioactive interferon, anti-(interferon alpha) serum, and bacterial adsorbent. The immune precipitation of the labeled interferon was inhibited by unlabeled interferon alpha, and 100 international reference units of interferon alpha could be measured in this way.
D000818:Animals; D002478:Cells, Cultured; D006801:Humans; D007369:Interferon Inducers / Q000494:pharmacology; D007372:Interferons / Q000096:biosynthesis*; D007930:Leucine / Q000378:metabolism; D008214:Lymphocytes / Q000378:metabolism*; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods; D013211:Staphylococcus aureus / Q000378:metabolism
D007369:Interferon Inducers; D007372:Interferons; D007930:Leucine