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Journal of Biomolecular Screening
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Development of an Immunofluorometric, High-Capacity, Cell-Based Assay for the Measurement of Human Type I and Type II Interferons

Lan Trinh

Department of Molecular Pharmacology, Berlex Biosciences Inc., Richmond, CA 94804

Russell Ziegler

Department of Cell Biology, Berlex Biosciences Inc., Richmond, CA 94804

Diane Watling

Imperial Cancer Research Fund, Lincoln's Inn Fields, London, WC2A 3PX UK

R. Michael Snider

Department of Molecular Pharmacology, Berlex Biosciences Inc., Richmond, CA 94804

Ed Croze

Department of Protein Biochemistry, Berlex Biosciences Inc., Richmond, CA 94804

We have developed a cell-based 96-well microtiter plate, high throughput assay for measuring both type I and type II interferon (IFN) activity on human cells. This assay makes use of a previously described IFN-specific reporter stably expressed in human HT 1080 cells. The induction of the reporter by IFN is determined by measuring the IFN-dependent expression of CD2 on the cell surface. The cytokine-induced expression of CD2 occurs within 48 h and is measured using a time-resolved fluorometric immunoassay. The limit of detection for type I IFN is >0.4 IU/ml. Interassay and intraassay coefficients of variation were 1.1% and 1.3% for the medium control (31 IU IFNß1b/ml), respectively. The limit of detection for type II IFN is >8 IU/ml, and the assay coefficients of variation are similar to those determined for type I IFNs. The level of sensitivity for this assay is comparable to other assays commonly used to measure IFN activity on cells. The current assay has an advantage over antiviral and antiproliferative assays, in that there is no requirement for the use of pathogenic virus or for determining viable cell numbers. The current assay is ideally suited for increasing sample screening and high-capacity automation, making it an excellent tool for drug discovery.

Journal of Biomolecular Screening, Vol. 4, No. 1, 33-37 (1999)
DOI: 10.1177/108705719900400106
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