Thrombolytic proteins profiling: High-throughput activity, selectivity, and resistance assays

Area of Science:

• Biochemistry
• Pharmacology
• Biotechnology

Background:

• Cardiovascular diseases and thrombotic events are leading causes of morbidity and mortality worldwide.
• Current thrombolytic enzyme therapies have suboptimal effectiveness and safety profiles.
• There is a need for improved thrombolytic agents or enhanced existing ones.

Purpose of the Study:

• To present optimized biochemical protocols for screening and assessing the therapeutic potential of thrombolytic biomolecules.
• To enable rapid and reliable evaluation of candidate thrombolytics' effectiveness, safety, and biological characteristics.
• To compare the properties of alteplase and tenecteplase using the developed methodology.

Main Methods:

• Development of optimized biochemical assays for thrombolytic biomolecule characterization.
• Assessment of enzymatic activity, fibrinolysis rate, fibrin/fibrinogen stimulation, fibrin selectivity, clot binding, and inhibition resistance.
• Application of the developed assays to evaluate alteplase and tenecteplase.

Main Results:

• The developed assays provide comprehensive data on multiple thrombolytic characteristics.
• Tenecteplase demonstrated increased fibrin selectivity and inhibition resistance compared to alteplase.
• These properties of tenecteplase correlate with its extended biological half-life.

Conclusions:

• The optimized biochemical protocols facilitate robust screening and therapeutic potential assessment of thrombolytic biomolecules.
• Tenecteplase may be a superior alternative to alteplase for thrombolytic treatment due to enhanced fibrin selectivity and inhibition resistance.
• The methodology aids in predicting a candidate's effectiveness, biological half-life, and potential side effects.