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- Title
A Neoepitope-Based Enzyme Immunoassay for Quantification of C1-Inhibitor in Complex with C1r and C1s.
- Authors
Fure, H.; Nlelsen, E. W.; Hack, C. E.; Mollnes, T. E.
- Abstract
Monoclonal antibodies (MoAb) recognizing neoepitopes exposed on activation products of complement proteins but hidden in the native components have been used for quantification of activated complement. A previously produced and characterized mouse MoAb, recognizing a neoepitope on the human plasma protein C1-inhibitor complexed with its substrates, was used to design an enzyme immunoassay for detection of C1-inhibitor complexed with C1r and C1s. These complexes are indicators of early classical complement pathway activation. The standard was serum activated with heat aggregated IgG defined to contain 1000 arbitrary units (AU)/ml. The lower detection limit was ≈0.05 AU/ml corresponding to 0.005% of fully activated serum. The reliability of the assay, including day-to-day variation, was tested. Intra-assay variation coefficients were 12% for low plasma control and 13% for high plasma control (<em>n</em> = 12 for both). Inter-assay variation coefficients were 12% for low control (<em>n</em>= 6), 19% for high control (<em>n</em> = 6) and 15% for the normal plasma control(<em>n</em> = 9). A 2.5-97.5 percentile reference range (normal blood donors) was 16-33 AU/ml. Two patients with systemic lupus erythematosus had considerably elevated plasma levels of the activation product (56 and 62 AU/ml). I and six patients with hereditary angioedema had normal plasma levels despite considerably reduced C1-inhibitor concentration. We conclude that the present method is sensitive and reliable for detection of early classical pathway activation and superior to previously published methods by utilizing neoepitope specificity and non-radiolabelled reagents.
- Subjects
IMMUNOASSAY; ENZYMES; MONOCLONAL antibodies; PROTEINS; SERUM
- Publication
Scandinavian Journal of Immunology, 1997, Vol 46, Issue 6, p553
- ISSN
0300-9475
- Publication type
Article
- DOI
10.1046/j.1365-3083.1997.d01-168.x