PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and Virology
Thymosin α1 (Tα1) is an immunostimulatory peptide for the treatment of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections and used as an immune enhancer, which also offers prospects in the context of COVID-19 infections and cancer. Manufacturing of this N-terminally acetylated 28-residue...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/1/124 |
| _version_ | 1797543578664173568 |
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| author | Uli Binder Arne Skerra |
| author_facet | Uli Binder Arne Skerra |
| author_sort | Uli Binder |
| collection | DOAJ |
| description | Thymosin α1 (Tα1) is an immunostimulatory peptide for the treatment of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections and used as an immune enhancer, which also offers prospects in the context of COVID-19 infections and cancer. Manufacturing of this N-terminally acetylated 28-residue peptide is demanding, and its short plasma half-life limits in vivo efficacy and requires frequent dosing. Here, we combined the PASylation technology with enzymatic in situ N-acetylation by RimJ to produce a long-acting version of Tα1 in <i>Escherichia coli</i> at high yield. ESI-MS analysis of the purified fusion protein indicated the expected composition without any signs of proteolysis. SEC analysis revealed a 10-fold expanded hydrodynamic volume resulting from the fusion with a conformationally disordered Pro/Ala/Ser (PAS) polypeptide of 600 residues. This size effect led to a plasma half-life in rats extended by more than a factor 8 compared to the original synthetic peptide due to retarded kidney filtration. Our study provides the basis for therapeutic development of a next generation thymosin α1 with prolonged circulation. Generally, the strategy of producing an N-terminally protected PASylated peptide solves three major problems of peptide drugs: (i) instability in the expression host, (ii) rapid degradation by serum exopeptidases, and (iii) low bioactivity because of fast renal clearance. |
| first_indexed | 2024-03-10T13:47:38Z |
| format | Article |
| id | doaj.art-8d8b3c6e6b7e4e9cb4643d4c2694feef |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T13:47:38Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-8d8b3c6e6b7e4e9cb4643d4c2694feef2023-11-21T02:28:56ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-12-0122112410.3390/ijms22010124PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and VirologyUli Binder0Arne Skerra1XL-protein GmbH, Lise-Meitner-Str. 30, 85354 Freising, GermanyLehrstuhl für Biologische Chemie, Technische Universität München, Emil-Erlenmeyer-Forum 5, 85354 Freising, GermanyThymosin α1 (Tα1) is an immunostimulatory peptide for the treatment of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections and used as an immune enhancer, which also offers prospects in the context of COVID-19 infections and cancer. Manufacturing of this N-terminally acetylated 28-residue peptide is demanding, and its short plasma half-life limits in vivo efficacy and requires frequent dosing. Here, we combined the PASylation technology with enzymatic in situ N-acetylation by RimJ to produce a long-acting version of Tα1 in <i>Escherichia coli</i> at high yield. ESI-MS analysis of the purified fusion protein indicated the expected composition without any signs of proteolysis. SEC analysis revealed a 10-fold expanded hydrodynamic volume resulting from the fusion with a conformationally disordered Pro/Ala/Ser (PAS) polypeptide of 600 residues. This size effect led to a plasma half-life in rats extended by more than a factor 8 compared to the original synthetic peptide due to retarded kidney filtration. Our study provides the basis for therapeutic development of a next generation thymosin α1 with prolonged circulation. Generally, the strategy of producing an N-terminally protected PASylated peptide solves three major problems of peptide drugs: (i) instability in the expression host, (ii) rapid degradation by serum exopeptidases, and (iii) low bioactivity because of fast renal clearance.https://www.mdpi.com/1422-0067/22/1/124biobettercancerCOVID-19drug deliveryhalf-life extensionimmunostimulatory peptide |
| spellingShingle | Uli Binder Arne Skerra PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and Virology International Journal of Molecular Sciences biobetter cancer COVID-19 drug delivery half-life extension immunostimulatory peptide |
| title | PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and Virology |
| title_full | PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and Virology |
| title_fullStr | PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and Virology |
| title_full_unstemmed | PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and Virology |
| title_short | PASylated Thymosin α1: A Long-Acting Immunostimulatory Peptide for Applications in Oncology and Virology |
| title_sort | pasylated thymosin α1 a long acting immunostimulatory peptide for applications in oncology and virology |
| topic | biobetter cancer COVID-19 drug delivery half-life extension immunostimulatory peptide |
| url | https://www.mdpi.com/1422-0067/22/1/124 |
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