Cell-free Synthesis of the Hirudin Variant 1 of the Blood-sucking Leech Hirudo Medicinalis

Overview

Journal Sci Rep

Specialty Science

Date 2020 Nov 14

PMID 33188246

Citations 6

Authors

Doreen A Wustenhagen

Phil Lukas

Christian Muller

Simone A Aubele

Jan-Peter Hildebrandt

Stefan Kubick

Affiliations

Soon will be listed here.

Abstract

Synthesis and purification of peptide drugs for medical applications is a challenging task. The leech-derived factor hirudin is in clinical use as an alternative to heparin in anticoagulatory therapies. So far, recombinant hirudin is mainly produced in bacterial or yeast expression systems. We describe the successful development and application of an alternative protocol for the synthesis of active hirudin based on a cell-free protein synthesis approach. Three different cell lysates were compared, and the effects of two different signal peptide sequences on the synthesis of mature hirudin were determined. The combination of K562 cell lysates and the endogenous wild-type signal peptide sequence was most effective. Cell-free synthesized hirudin showed a considerably higher anti-thrombin activity compared to recombinant hirudin produced in bacterial cells.

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