Details
| Originalsprache | Englisch |
|---|---|
| Aufsatznummer | 39 |
| Fachzeitschrift | Journal of Functional Biomaterials |
| Jahrgang | 10 |
| Ausgabenummer | 3 |
| Frühes Online-Datum | 2 Sept. 2019 |
| Publikationsstatus | Veröffentlicht - Sept. 2019 |
Abstract
Proteins derived from the natural extracellular matrix like collagen or gelatin are common in clinical research, where they are prized for their biocompatibility and bioactivity. Cells are able to adhere, grow and remodel scaffolds based on these materials. Usually, collagen and gelatin are sourced from animal material, risking pathogenic transmission and inconsistent batch-to-batch product quality. A recombinant production in yeast circumvents these disadvantages by ensuring production with a reproducible quality in animal-component-free media. A gelatin mimetic protein, based on the alpha chain of human collagen I, was cloned in Pichia pastoris under the control of the methanol-inducible alcohol oxidase (AOX1) promoter. A producing clone was selected and cultivated at the 30 L scale. The protein was secreted into the cultivation medium and the final yield was 3.4 g·L−1. Purification of the target was performed directly from the cell-free medium by size exclusion chromatography. The gelatin mimetic protein was tested in cell culture for biocompatibility and for promoting cell adhesion. It supported cell growth and its performance was indistinguishable from animal-derived gelatin. The gelatin-mimetic protein represents a swift strategy to produce recombinant and human-based extracellular matrix proteins for various biomedical applications.
ASJC Scopus Sachgebiete
- Werkstoffwissenschaften (insg.)
- Biomaterialien
- Ingenieurwesen (insg.)
- Biomedizintechnik
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in: Journal of Functional Biomaterials, Jahrgang 10, Nr. 3, 39, 09.2019.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Production of a Recombinant Non-Hydroxylated Gelatin Mimetic in Pichia pastoris for Biomedical Applications
AU - Gellermann, Pia
AU - Schneider-Barthold, Caroline
AU - Bolten, Svenja Nicolin
AU - Overfelt, Ethan
AU - Scheper, Thomas
AU - Pepelanova, Iliyana
N1 - Funding information: Acknowledgments: This work was performed within the framework of the BIOFABRICATION FOR NIFE initiative, financially supported by the Ministry of Science and Culture (MWK) of Lower Saxony, Germany. We would like to thank Beate Jaschok-Kentner from the Helmholtz Zentrum (HZI) Braunschweig for performing the Edman sequence analysis. The publication of this article was funded by the Open Access fund of Leibniz Universität Hannover. Funding: This research was funded by the BIOFABRICATION FOR NIFE initiative, financially supported by the Ministry of Science and Culture (MWK) of Lower Saxony, Germany.
PY - 2019/9
Y1 - 2019/9
N2 - Proteins derived from the natural extracellular matrix like collagen or gelatin are common in clinical research, where they are prized for their biocompatibility and bioactivity. Cells are able to adhere, grow and remodel scaffolds based on these materials. Usually, collagen and gelatin are sourced from animal material, risking pathogenic transmission and inconsistent batch-to-batch product quality. A recombinant production in yeast circumvents these disadvantages by ensuring production with a reproducible quality in animal-component-free media. A gelatin mimetic protein, based on the alpha chain of human collagen I, was cloned in Pichia pastoris under the control of the methanol-inducible alcohol oxidase (AOX1) promoter. A producing clone was selected and cultivated at the 30 L scale. The protein was secreted into the cultivation medium and the final yield was 3.4 g·L−1. Purification of the target was performed directly from the cell-free medium by size exclusion chromatography. The gelatin mimetic protein was tested in cell culture for biocompatibility and for promoting cell adhesion. It supported cell growth and its performance was indistinguishable from animal-derived gelatin. The gelatin-mimetic protein represents a swift strategy to produce recombinant and human-based extracellular matrix proteins for various biomedical applications.
AB - Proteins derived from the natural extracellular matrix like collagen or gelatin are common in clinical research, where they are prized for their biocompatibility and bioactivity. Cells are able to adhere, grow and remodel scaffolds based on these materials. Usually, collagen and gelatin are sourced from animal material, risking pathogenic transmission and inconsistent batch-to-batch product quality. A recombinant production in yeast circumvents these disadvantages by ensuring production with a reproducible quality in animal-component-free media. A gelatin mimetic protein, based on the alpha chain of human collagen I, was cloned in Pichia pastoris under the control of the methanol-inducible alcohol oxidase (AOX1) promoter. A producing clone was selected and cultivated at the 30 L scale. The protein was secreted into the cultivation medium and the final yield was 3.4 g·L−1. Purification of the target was performed directly from the cell-free medium by size exclusion chromatography. The gelatin mimetic protein was tested in cell culture for biocompatibility and for promoting cell adhesion. It supported cell growth and its performance was indistinguishable from animal-derived gelatin. The gelatin-mimetic protein represents a swift strategy to produce recombinant and human-based extracellular matrix proteins for various biomedical applications.
KW - Gelatin-mimetic protein
KW - Komagataella pfaffi (Pichia pastoris)
KW - Recombinant ECM-protein
KW - Recombinant gelatin
UR - http://www.scopus.com/inward/record.url?scp=85073074128&partnerID=8YFLogxK
U2 - 10.3390/jfb10030039
DO - 10.3390/jfb10030039
M3 - Article
AN - SCOPUS:85073074128
VL - 10
JO - Journal of Functional Biomaterials
JF - Journal of Functional Biomaterials
IS - 3
M1 - 39
ER -