A simple methodology for the routine production and partial purification of human lymphoblastoid interferon.
We report a methodology suitable for the large scale production and partial purification of human lymphoblastoid interferon with a minimum expense and reasonable (10%) degree of purity of product. The cells used were Namalwa lymphoblastoid cells, which have the advantage of being both easy to grow and well characterized. They were grown in RPMI 1640 containing 10% fetal calf serum to a density of 1.5 - 2 X 10(6) cells/ml and then diluted 50% by the addition of serum free medium containing 2mM sodium butyrate. After 48 hours, the medium was removed and the cells induced to produce interferon by Sendai virus. Typical initial interferon titres were in the region of 4 - 4.8 log units/ml, while specific activities were in the region of 10(6) units/mg protein. The initial stage in the purification involved batchwise adsorption of the interferon to Procion red- HE7B Sepharose CL6B . The Sepharose was then packed into a column, washed with 0.5M KC1 and the interferon eluted with 2M KC1. The interferon was further purified by gel filtration to give an activity of approximately 10(7) units/mg protein. Yields were between 30-50% of the initial interferon in a volume of 25 ml. Further separation of the components of the heterogeneous alpha interferon could be obtained on Procion blue-HER Sepharose, or by utilizing reverse phase HPLC.
D000818:Animals; D002051:Burkitt Lymphoma; D002460:Cell Line; D002522:Chlorocebus aethiops; D006801:Humans; D007370:Interferon Type I / Q000096:biosynthesis* / Q000302:isolation & purification / Q000633:toxicity; D007668:Kidney; D010222:Parainfluenza Virus 1, Human; D012672:Semliki forest virus / Q000187:drug effects
D007370:Interferon Type I