A novel biosensor for measuring plasmin activity

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Plasmin is crucial for fibrinolysis, degrading fibrin clots.
• Accurate measurement of plasmin generation is essential for preclinical and clinical research.
• Current methods for assessing fibrinolysis require improvement.

Purpose of the Study:

• To design, synthesize, and evaluate a novel fluorescence resonance energy quenching-based plasmin sensor (FPS) protein.
• To establish a reliable method for monitoring plasmin generation.
• To assess the FPS protein's efficiency and specificity in detecting plasmin.

Main Methods:

• Developed and synthesized a novel FPS protein utilizing fluorescence resonance energy quenching.
• Monitored FPS hydrolysis via loss of fluorescence resonance energy quenching to assess efficiency and specificity.
• Employed plasma-based plasmin generation assays to evaluate fibrinolytic cofactors and inhibitors.

Main Results:

• The FPS protein exhibited higher efficiency for plasmin compared to peptide sensors and was unaffected by plasmin inhibited by α2-macroglobulin.
• FPS successfully measured plasmin generation in clotting plasma, demonstrating sensitivity to fibrinolytic modulators like α2-antiplasmin, aprotinin, tranexamic acid, and fibrin.
• Plasmin generation measurements using FPS in healthy volunteers showed consistent results with limited variation.

Conclusions:

• The developed FPS protein offers a promising tool for evaluating fibrinolysis in both preclinical and clinical settings.
• FPS can be utilized to measure plasmin generation, aiding in the study of fibrinolytic disorders.
• This sensor provides a reliable and consistent method for assessing plasmin activity, potentially improving diagnostic and therapeutic strategies.