An in vitro bioassay for quantitation of human interferons by measurements of antiproliferative activity on a continuous human lymphoma cell line.
Interferons have, in addition to their antiviral effects, been shown to possess several non-antiviral activities. In this study, an in vitro bioassay for interferon alpha (IFN-alpha) preparations based on their antiproliferative effect in cultured Daudi cells has been developed. Briefly, about 10(5) cells per ml treated with different concentrations of IFN were incubated under standard culture conditions for 3 days. Two different end points, i.e. incorporation of [3H]thymidine and final cell density, were used and responses were evaluated according to established pharmacopoeial principles for quantification of biomolecules. Both methods gave similar results. However, measurement of final cell density yielded the most precise results. The proposed assay, with an effective assay range of 1-10 IU/ml (approximately 0.2-2 x 10(-12)M, had a high sensitivity and precision as well as a good reproducibility. Compared with antiviral assays, it is less resource demanding. In conclusion, the in vitro bioassay described is well suited for potency determinations of IFN-alpha and probably also IFN-beta preparations.
Nederman
T
T
National Board of Health and Welfare, Department of Drugs, Uppsala, Sweden.
Karlström
E
E
Sjödin
L
L
D001681:Biological Assay; D002455:Cell Division / Q000187:drug effects; D006801:Humans; D007370:Interferon Type I / Q000032:analysis* / Q000494:pharmacology; D008223:Lymphoma; D011994:Recombinant Proteins; D012015:Reference Standards; D013936:Thymidine / Q000378:metabolism; D014407:Tumor Cells, Cultured
D007370:Interferon Type I; D011994:Recombinant Proteins; D013936:Thymidine