Monoclonal antibodies to human interleukin-1 beta and their use in a sensitive two-site enzyme linked immunosorbent assay.
Five Monoclonal Antibodies (MAbs) coded respectively #111, 122, 206, 209 and 609 were produced against human recombinant Interleukin-1 beta (rIL-1 beta, amino acids 117-269). Four of these MAbs (#111, 122, 206 and 609) have been identified able to inhibit the biological activity of recombinant and natural Interleukin-1 beta. Competition studies suggested that three non-overlapping epitopes of the molecule were recognized by the MAbs. MAbs #609 and 206 have been selected on the basis of high affinity and used together in a two-site ELISA to detect IL-1 beta. The sensitivity of this ELISA was 1 pg/well (10-20 pg/ml). The assay did not detect rHuIL-1 alpha, rHuIL-2, rHuTNF-alpha, rHuIFN-gamma, rHuIL-6, and natural a- and b-FGF. The immunoassay was then compared to current assay such as co-mitogenic effect on murine thymocytes for detection of IL-1 beta in the intra- and extracellular compartments of IFN-gamma and/or LPS-stimulated human blood monocytes. We confirm that IFN-gamma potentiates IL-1 beta secretion in response to LPS (even with high dose 1 microgram/ml) but not the intracellular precursor of IL-1 beta. This immunoassay could be used also to detect IL-1 beta in plasma, sera and synovial fluids.
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Département de Biotechnologies, Roussel-Uclaf, Romainville, France.
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D000818:Animals; D000911:Antibodies, Monoclonal; D001826:Body Fluids / Q000032:analysis; D002455:Cell Division; D003124:Colorimetry; D015232:Dinoprostone / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007118:Immunoassay; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000032:analysis / Q000276:immunology*; D007526:Isoelectric Point; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008934:Mitogens; D009000:Monocytes / Q000187:drug effects; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D013950:Thymus Gland / Q000166:cytology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007375:Interleukin-1; D008070:Lipopolysaccharides; D008934:Mitogens; D011994:Recombinant Proteins; D007371:Interferon-gamma; D015232:Dinoprostone