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Extracellular Vesicle Tissue Factor Activity Assay
Published on: December 29, 2023
A simplified and high-throughput chromogenic assay for testing tissue factor-dependent procoagulant activity
Wenwen Jiang1, Junping Kou, Shengtao Yuan
1Department of Complex Prescription of TCM, School of Chinese Material Medicine, China Pharmaceutical University, Nanjing, China.
Journal of Biomolecular Screening
|February 12, 2011
Summary
A new, simple assay effectively screens for tissue factor (TF) inhibitors, crucial for treating cardiovascular diseases. This high-throughput method utilizes readily available components, making TF inhibitor discovery more accessible.
Area of Science:
- Biochemistry
- Pharmacology
- Assay Development
Background:
- Tissue factor (TF) initiates coagulation and is a therapeutic target for cardiovascular diseases.
- Existing assays for TF activity are often complex and costly.
- Selective small-molecule TF inhibitors are scarce.
Purpose of the Study:
- To develop a simple, high-throughput chromogenic assay for screening TF inhibitors.
- To validate the assay's sensitivity and precision compared to existing methods.
- To identify potential new TF inhibitors using the developed assay.
Main Methods:
- A one-stage chromogenic assay using commercial human prothrombin complex.
- Incubation of cell lysates with prothrombin complex, followed by addition of chromogenic substrate Xa.
- Absorbance measurement at 405 nm in 96-well or 384-well plates.
Main Results:
- The simplified assay demonstrated high sensitivity and precision for measuring TF procoagulant activities.
- Known TF inhibitors (curcumin, simvastatin) were validated using the assay.
- Berberine and cryptotanshinone showed suppressed TF activity in human monocytes, indicating potential as new inhibitors.
Conclusions:
- The developed chromogenic assay is a simple, efficient, and high-throughput method for TF inhibitor screening.
- This assay facilitates the discovery of novel TF inhibitors for potential therapeutic applications.
- Berberine and cryptotanshinone warrant further investigation as potential TF-targeted therapeutics.
