PMID: 2494265Mar 31, 1989Paper

Single step screening of monoclonal antibodies against interferon-gamma-induced surface molecules on human monocytes

Journal of Immunological Methods
B A VanceP M Guyre

Abstract

Interferon-gamma (IFN-gamma) activation of human monocytes in vitro results in enhanced phagocytosis and cellular cytotoxicity. These enhanced effector functions are attributable, at least in part, to increased expression of recognition molecules on the plasma membrane. In this article we report a rapid screening procedure for the primary selection of monoclonal antibodies (mAbs) which bind to cell surface molecules, the expression of which is increased or decreased by IFN-gamma. The procedure is based on flow cytometric analysis of mixed cell populations. Mouse mAbs were prepared using human monocytes cultured for 40 h with 400 U/ml of IFN-gamma as the immunogen. The hybridoma supernatants were screened using mixtures of six cell populations, some of which were pretreated with IFN-gamma for 40 h. Cells included in the mixture were chosen for their distinctive light scatter profiles. mAbs of interest were identified by preferential binding to monocytes and increased or decreased binding to monocytes treated with IFN-gamma. This procedure allowed us to screen several hundred clones per day, and to immediately eliminate mAbs that bound to B cells, T cells, neutrophils, and several cell lines. We selected ten mAbs which bound to s...Continue Reading

References

Sep 1, 1987·European Journal of Immunology·G D KeizerJ E De Vries
Sep 1, 1986·Cellular Immunology·S J MentzerD V Faller
Dec 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·F Sanchez-MadridT A Springer
Jan 1, 1982·Journal of Immunological Methods·M R Loken, A M Stall

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Citations

Jan 1, 1989·Current Opinion in Immunology·P M Lydyard
May 1, 1997·Veterinary Immunology and Immunopathology·P E JollyW J Sapp
Dec 18, 2004·Journal of Immunological Methods·Annet van Royen-KerkhofJan G J van de Winkel

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