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Updated: Jan 6, 2026
Author Spotlight: Advancing Thrombolytic Testing by Integrating Flow Dynamics in In Vitro Models
Published on: April 19, 2024
Thrombolytic proteins profiling: High-throughput activity, selectivity, and resistance assays
Martin Toul1,2, Alan Strunga1,2, Jiri Damborsky1,2
1Loschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk University, Brno, Czech Republic.
Insights
Optimized biochemical assays enable robust screening of clot-dissolving thrombolytic biomolecules. Tenecteplase shows superior fibrin selectivity and inhibition resistance compared to alteplase for treating cardiovascular diseases.
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.
Abstract:
Cardiovascular diseases, including thrombotic events such as ischemic stroke, pulmonary embolism, and myocardial infarction, are among the leading causes of morbidity and disability worldwide. The application of clot-dissolving thrombolytic enzymes is a cost-effective therapeutic intervention for these life-threatening conditions. However, the effectiveness and safety profiles of current drugs are suboptimal, necessitating the discovery of new medicines or the engineering and enhancement of the existing ones. Here, we present a set of optimized biochemical protocols that allow robust screening and the therapeutic potential assessment of thrombolytic biomolecules. The assays provide information on multiple characteristics such as enzymatic activity, fibrinolysis rate, fibrin and fibrinogen stimulation, fibrin selectivity, clot binding affinity, and inhibition resistance. Such detailed characterization enables a rapid and reliable evaluation of candidate effectiveness and provides an indication of biological half-life, associated bleeding complications, and other side effects. We demonstrate the credibility of the methodology by applying it to the two most widely used thrombolytic drugs: alteplase (Activase®/Actilyse®) and tenecteplase (Metalyse®/TNKase®). Consistent with previous studies, tenecteplase exhibited increased fibrin selectivity and inhibition resistance, which explains its extended half-life. Our findings reinforce the growing consensus that tenecteplase may be a superior alternative to alteplase for thrombolytic treatment.
