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Heparin-Bead Extraction Enhanced Fluorogenic Aptamer-Thrombin Composite Reporter Enables Sensitive and Rapid
He Yan1, Zimu Tian1, Emma Ziegenbein2
1Department of Chemistry, University of Florida, Gainesville, Florida 32611, United States.
ACS Sensors
|August 23, 2024
Summary
Antithrombin (AT) deficiency impacts patients during cardiac surgery and critical illness. A new HExGATOR method rapidly and accurately detects functional AT in plasma, improving patient monitoring and personalized anticoagulant therapy.
Area of Science:
- Biochemistry
- Medical Diagnostics
- Biotechnology
Background:
- Antithrombin (AT) deficiency is critical in extracorporeal circulation during cardiac surgery, leading to inflammation and vascular damage.
- AT levels decrease in conditions like sepsis, major trauma, ECMO, and eclampsia, necessitating precise monitoring.
- Current chromogenic and ELISA assays for AT lack real-time reliability and face interference issues, creating a clinical monitoring gap.
Purpose of the Study:
- To develop a rapid, sensitive, and precise method for detecting functional antithrombin (AT) in plasma.
- To address the limitations of traditional AT detection methods in clinical settings.
- To facilitate accurate AT restoration and personalized anticoagulant therapy.
Main Methods:
- Development of a heparin-bead extraction enhanced fluoroGenic aptamer-thrombin composite reporter (HExGATOR).
- Utilized thrombin-binding aptamers and a fluorescence "turn on" mechanism for signal generation upon AT interaction.
- Employed heparin-bead solid-phase extraction to minimize plasma-derived signal-background interference.
Main Results:
- Achieved highly sensitive and rapid detection of functional AT in plasma within 5 to 20 minutes.
- Established a limit of detection of 15.11 nM for AT.
- Demonstrated significant reduction in signal-background interference using the heparin-bead extraction method.
Conclusions:
- The HExGATOR assay offers a promising alternative to traditional AT detection methods.
- This novel approach enables rapid and precise monitoring of functional AT in clinical settings.
- Facilitates personalized anticoagulant therapy through accurate AT level assessment.
