title: A new enzyme-linked immunosorbent assay recognizing both rat and human activated coagulation Factor XII (FXIIa).
abstract: We describe the first sandwich enzyme-linked immunosorbent assay (ELISA) capable of recognizing both rat and human activated coagulation Factor XII (FXIIa). Increased plasma concentrations of FXIIa have been associated with adverse outcomes in several cardiovascular disease states. In humans, the FXIIa antigen in plasma is quantified by a direct sandwich ELISA employing an antibody (mAb 2/215) raised against its β-fragment (β-FXIIa), but this assay is unable to detect rat β-FXIIa antigen. Thus, experimental models are at present limited in their capacity to reveal mechanisms by which FXIIa might contribute to cardiovascular pathology. Consistent with overlap between human and rat FXIIa protein epitope sequences, Western blot analysis and ELISA demonstrate that another previously developed antibody against human FXIIa (mAb 201/9) detects β-FXIIa in both human and rat plasma, with no evidence for cross-reactivity with the FXII zymogen or FXIIa complexed with its endogenous inhibitor. The mAb 201/9 based ELISA identified similar elevations in FXIIa in plasma from rats and humans with chronic renal failure. The capacity of this ELISA to identify rat plasma FXIIa has potential application to a wide range of experimental models of human cardiovascular disease.
authors.affiliation: Heart and Stroke/Richard Lewar Centre of Excellence, University of Toronto, Toronto, ON, Canada.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Papageorgiou
authors.forename: Erik L
authors.initials: EL
authors.lastname: Yeo
authors.forename: Peter H
authors.initials: PH
authors.lastname: Backx
authors.forename: John S
authors.initials: JS
authors.lastname: Floras
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D002318:Cardiovascular Diseases / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology; D015956:Factor XIIa / Q000032:analysis / Q000276:immunology*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D015956:Factor XIIa