Establishment of dye-uptake method (A375 assay) for quantitative measurement of IL-1: correlation with LAF assay.
The dye-uptake method (A375 assay) for quantitative measurement of biological activity or recombinant human interleukin-1 (IL-1) was established based on the cytostatic activity of IL-1 on human melanoma A375 cells. Although various factors, e.g., incubation temperature, inoculum cell size and cell growth phase, were found to influence the titration to some degree, the assay-to-assay fluctuation in the observed titers determined by this method could be made minimum after normalization with the observed titer of a standard IL-1. A good correlation was observed between A375 and LAF assays. Thus, A375 assay was shown to be a stable and reliable method and could be replaced with LAF assay for quantitative measurement of recombinant human IL-1.
Nakano
K
K
Research Laboratories, Dainippon Pharmaceutical Co., Ltd., Suita-Osaka, Japan.
Okugawa
K
K
Hayashi
H
H
Abe
S
S
Sohmura
Y
Y
Tsuboi
T
T
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D007375:Interleukin-1 / Q000032:analysis* / Q000494:pharmacology; D008213:Lymphocyte Activation; D008545:Melanoma; D008722:Methods; D008751:Methylene Blue; D051379:Mice; D008809:Mice, Inbred C3H; D011994:Recombinant Proteins / Q000032:analysis* / Q000494:pharmacology; D013053:Spectrophotometry / Q000379:methods; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology
D007375:Interleukin-1; D011994:Recombinant Proteins; D008751:Methylene Blue