Cell-free synthesis of the hirudin variant 1 of the blood-sucking leech Hirudo medicinalis
- 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.
| Author details: | Doreen Anja Wüstenhagen, Phil LukasORCiDGND, Christian Müller, Simone A. Aubele, Jan-Peter HildebrandtORCiDGND, Stefan KubickORCiD |
|---|---|
| DOI: | https://doi.org/10.1038/s41598-020-76715-w |
| ISSN: | 2045-2322 |
| Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/33188246 |
| Title of parent work (English): | Scientific reports |
| Publisher: | Macmillan Publishers Limited, part of Springer Nature |
| Place of publishing: | London |
| Publication type: | Article |
| Language: | English |
| Date of first publication: | 2020/11/13 |
| Publication year: | 2020 |
| Release date: | 2023/09/19 |
| Volume: | 10 |
| Issue: | 1 |
| Article number: | 19818 |
| Number of pages: | 13 |
| Funding institution: | Projekt DEAL |
| Organizational units: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie |
| DDC classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
| Peer review: | Referiert |
| Publishing method: | Open Access / Gold Open-Access |
| License (German): |  CC-BY - Namensnennung 4.0 International |
