A sensitive immunochemical assay for measuring the concentration of the activated protein C-protein C inhibitor complex in plasma: Use of a catcher antibody specific for the complexed/cleaved form of the inhibitor

Citation
K. Strandberg et al., A sensitive immunochemical assay for measuring the concentration of the activated protein C-protein C inhibitor complex in plasma: Use of a catcher antibody specific for the complexed/cleaved form of the inhibitor, THROMB HAEM, 86(2), 2001, pp. 604-610
Citations number
37
Categorie Soggetti
Cardiovascular & Hematology Research
Journal title
THROMBOSIS AND HAEMOSTASIS
ISSN journal
03406245 → ACNP
Volume
86
Issue
2
Year of publication
2001
Pages
604 - 610
Database
ISI
SICI code
0340-6245(200108)86:2<604:ASIAFM>2.0.ZU;2-H
Abstract
Activated protein C (APC) is a serine proteinase that regulates blood coagu lation. In plasma it is inhibited mainly by the protein C inhibitor (PCI). The plasma concentrations of APC-PCI complex is increased in hypercoagulati ve states such as deep venous thrombosis. Formation of the APC-PCI complex induces a drastic conformational change in PCI that exposes new epitopes (n eoepitopes) on the molecule. We have devised a simple immunofluorometric sa ndwich assay for measurements of the concentrations of APC-PCI complex, emp loying as the catcher, a monoclonal antibody that has a high affinity (K-D approximate to 4 X 10(-11) M) for a complexation-specific neoepitope that i s expressed on PCI. A monoclonal antibody against protein C is employed as the tracer. The method gives a linear dose-response curve (0.06-50 mug/l), has a low detection limit (0.06 mug/l) and no crossreactivity with native P CI at physiologic plasma concentrations. We have now determined the concent ration of the APC-PCl complex in healthy individuals.