Quantitative measurement of human interleukin 2 receptor levels with intact and detergent-solubilized human T-cells |
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| Authors: | Steven K. Dower Steven H. Hefeneider Alan R. Alpert David L. Urdal |
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| Affiliation: | Immunex Corporation, 51 University St., Seattle, WA 98105, U.S.A. |
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| Abstract: | ![]()
2A3 monoclonal antibody (γ1, κ) is a novel high-affinity reagent for detecting the human interleukin 2 (IL-2) receptor. The antibody inhibits IL-2 binding to its receptor and is an antagonist of IL-2 action. Detailed analysis of the mechanism of binding of the IgG, (Fab')2 and Fab' of 2A3 antibody shows that the bivalent species cross-link on the cell surface when bound. Measurements of IL-2 receptor expression on digitonin-permeabilized cells suggest that the intracellular pool of receptors is small. The antibody will bind to IL-2 receptors on glutaraldehyde-fixed cells in the presence of Triton X-100. This property is used in designing an assay for quantitative measurements of IL-2 receptor concn in solution. This assay can be used to monitor receptor protein during purification to homogeneity. |
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| Keywords: | PHA phytohemagglutinin PBL peripheral blood leukocytes ELISA enzyme-linked immunoabsorbent assay HAT hypoxanthine, aminopterin, thymidine BSA bovine serum albumin PBS PBS-Triton PBS containing 1% Triton X-100 (w/v) binding medium IL-2 interleukin 2 TCGF T-cell growth factor IODOGEN 1,3,4,6-tetrachloro-3α,6α-diphenylglycouril |
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