A new enzyme-linked immunosorbent assay recognizing both rat and human activated coagulation Factor XII (FXIIa)

Area of Science:

• Biochemistry
• Immunology
• Cardiovascular Research

Background:

• Elevated plasma activated coagulation Factor XII (FXIIa) is linked to cardiovascular disease.
• Existing assays cannot measure rat FXIIa, limiting preclinical research.
• Understanding FXIIa's role in cardiovascular pathology requires cross-species measurement capabilities.

Purpose of the Study:

• To develop the first sandwich ELISA for quantifying both human and rat FXIIa.
• To enable the use of experimental models for studying FXIIa in cardiovascular disease.

Main Methods:

• Developed a novel sandwich ELISA using a monoclonal antibody (mAb 201/9) that recognizes a common epitope on human and rat FXIIa.
• Validated the assay using Western blot and ELISA to confirm specificity for FXIIa and rule out cross-reactivity with FXII zymogen or complexes.
• Measured FXIIa levels in plasma samples from humans and rats with chronic renal failure.

Main Results:

• The new ELISA successfully detects activated coagulation Factor XII (FXIIa) in both human and rat plasma.
• The assay shows no cross-reactivity with the FXII zymogen or FXIIa bound to its inhibitor.
• Similar elevations in FXIIa were observed in plasma from rats and humans with chronic renal failure.

Conclusions:

• A novel, specific ELISA for measuring FXIIa in both human and rat plasma has been established.
• This assay overcomes previous limitations, allowing for the study of FXIIa in diverse experimental models.
• The findings support the potential application of this ELISA in a wide range of cardiovascular disease research.