ISSN:
1365-3083
Source:
Blackwell Publishing Journal Backfiles 1879-2005
Topics:
Medicine
Notes:
A synthetic peptide of human recombinant interleukin 1β (hrIL-1β) 165–186, which exhibits biological activity in the human fibroblast assay, was used as an immunizing antigen to obtain a murine monoclonal antibody (MoAb) termed FIB 1 This MoAb, an IgG1, reacts specifically with hrIL-1β. but not with hrIL-1α, as measured in enzyme-linked immunosorbent assays (ELISA). The MoAb FIB 1 detects the characteristic 17 kDa IL-1 protein in Western blots. Binding to the antigen is specific, as deduced also from the close correlation of ELISA immunoreactivity with IL-1 biological activity. The anti-IL-1β 165–186 Ab specifically neutralizes the biological activity of hrIL-1β and native IL-1, as measured by the IL-1-induced proliferation of murine thymocytes and human fibroblasts and the IL-1-dependent IL-2 production by murine T cells (EL4-6.1). Fifty per cent of hrIL-1β activity (25 U/ml, or 0.25 ng/ml) has neutralized by less than 30μg/ml of MoAb. Furthermore, FIB 1 recognizes intracellular IL-1 in lipopolysaccharide-stimulated human peripheral blood mononuclear cells. The anti-IL-1β 165–186 Ab does not react with the shorter IL-1β fragment 161–173 in solid-phase ELISA, therefore the binding region seems to be localized in the amino acid sequence VALGLKEKN-LYLS. A sandwich-ELISA, using a polyclonal sheep anti-IL-1β 251–269 Ab as the capture antibody and anti-1β-165–186 MoAb as the detecting probe, allowed the determination of IL-1β from crude culture supernatants.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1111/j.1365-3083.1989.tb02462.x
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