A novel ELISA for mouse activated protein C in plasma

José A. Fernández, Steven R. Lentz, Denis M Dwyre, John H. Griffin

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

The Protein C pathway plays a crucial role in the regulation of thrombosis and inflammation. One of the tools that researchers presently use to elucidate mechanisms of action of activated protein C (APC) is the use of transgenic or gene deletion murine models. To correlate observations in these murine models with the APC levels, there is a need for a sensitive and specific assay for circulating murine APC in plasma. We developed an immunological assay to measure the physiological and pharmacologic levels of circulating murine APC. The sandwich ELISA uses an anti-murine anti-protein C antibody capture antibody and human protein C inhibitor (PCI) as a detection reagent, taking advantage of the facts that the mouse lacks plasma PCI and that human PCI forms a 1/1 stable complex with mouse APC. The amount of complex APC:PCI is detected with an anti-human PCI monoclonal antibody. The assay shows improved sensitivity versus enzyme immunocapture assays commonly used to detect human APC and considerably reduces the processing time.

Original languageEnglish (US)
Pages (from-to)174-181
Number of pages8
JournalJournal of Immunological Methods
Volume314
Issue number1-2
DOIs
StatePublished - Jul 31 2006
Externally publishedYes

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Keywords

  • Activated protein C
  • ELISA
  • Mouse
  • Protein C
  • Protein C inhibitor

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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