WEDGE: An anticoagulant thrombin mutant produced by autoactivation

D. C. Wood, L. A. Pelc, N. Pozzi, M. Wallisch, N. G. Verbout, E. I. Tucker, A. Gruber, Enrico Di Cera

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Summary: Background: The production of therapeutically relevant proteases typically involves activation of a zymogen precursor by external enzymes, which may raise regulatory issues about availability and purity. Recent studies of thrombin precursors have shown how to engineer constructs that spontaneously convert to the mature protease by autoactivation, without the need for external enzymes. Objectives: Autoactivation is an innovative strategy that promises to simplify the production of proteases of therapeutic relevance, but has not been tested in practical applications. The aim of this study was to provide a direct test of this strategy. Methods: An autoactivating version of the thrombin mutant W215A/E217A (WE), which is currently in preclinical development as an anticoagulant, was engineered. Results and Conclusions: The autoactivating version of WE can be produced in large quantities, like WE made in BHK cells or Escherichia coli, and retains all significant functional properties in vitro and in vivo. The results serve as proof of principle that autoactivation is an innovative and effective strategy for the production of trypsin-like proteases of therapeutic relevance.

Original languageEnglish (US)
Pages (from-to)111-114
Number of pages4
JournalJournal of Thrombosis and Haemostasis
Volume13
Issue number1
DOIs
StatePublished - Jan 1 2015

Keywords

  • Anticoagulants
  • Blood coagulation factors
  • Protein engineering
  • Thrombin
  • Zymogens

ASJC Scopus subject areas

  • Hematology

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    Wood, D. C., Pelc, L. A., Pozzi, N., Wallisch, M., Verbout, N. G., Tucker, E. I., Gruber, A., & Di Cera, E. (2015). WEDGE: An anticoagulant thrombin mutant produced by autoactivation. Journal of Thrombosis and Haemostasis, 13(1), 111-114. https://doi.org/10.1111/jth.12774