Abstract
Objective
To compare the effects of human Trypsin-1 signal peptide and pro-peptide on the expression and secretion efficiency of human Interleukin-25 from mammalian cells.
Results
The signal peptide and combined signal peptide-pro-peptide sequence of human Trypsin-1 improved the secretion of human IL-25 from 1.7 to 3.2 µg/ml and 1.7 to 8.2 µg/ml, respectively. Deletion analysis identified the minimal Trypsin-1 derived secretion domain that maintains improved human Interleukin-25 production and secretion. The presence of Trypsin-1 pro-peptide sequence does not affect the function of secreted human Interleukin-25.
Conclusion
The Trypsin-1 signal peptide-pro-peptide sequence increased human IL-25 expression and secretion in mammalian cells by fivefold.
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Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We would like to thanks members of the Protein Technology Group for scientific discussions.
Supplementary Information
Supplementary Fig. 1 — Amino acid sequences for recombinant hIL25-His tagged protein constructs used in this study.
Supplementary Fig. 2 — Amino acid sequences for recombinant human IL-25-His N-terminal and C-terminal deletion constructs used in this study.
Supplementary Fig. 3 — Amino acid sequences for recombinant human IL25-His containing the native IL25 signal peptide with the C-terminal ∆C3 sequence.
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by MM and CW. The first draft of the manuscript was written by MM and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Mullin, M.J., Wilkinson, C., Hiles, I. et al. Improved secretion of recombinant human IL-25 in HEK293 cells using a signal peptide-pro-peptide domain derived from Trypsin-1. Biotechnol Lett 43, 757–765 (2021). https://doi.org/10.1007/s10529-020-03072-z
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DOI: https://doi.org/10.1007/s10529-020-03072-z