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Recombinant self-assembling peptides as biomaterials for tissue engineering
Synthetic nanostructures based on self-assembling systems that aim to mimic natural extracellular matrix are now being used as substrates in tissue engineering applications. Peptides are excellent starting materials for the self-assembly process as they can be readily synthesised both chemically and...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier Science
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3111806/ https://www.ncbi.nlm.nih.gov/pubmed/20932572 http://dx.doi.org/10.1016/j.biomaterials.2010.08.051 |
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author | Kyle, Stuart Aggeli, Amalia Ingham, Eileen McPherson, Michael J. |
author_facet | Kyle, Stuart Aggeli, Amalia Ingham, Eileen McPherson, Michael J. |
author_sort | Kyle, Stuart |
collection | PubMed |
description | Synthetic nanostructures based on self-assembling systems that aim to mimic natural extracellular matrix are now being used as substrates in tissue engineering applications. Peptides are excellent starting materials for the self-assembly process as they can be readily synthesised both chemically and biologically. P(11)-4 is an 11 amino acid peptide that undergoes triggered self-assembly to form a self-supporting hydrogel. It exists as unimers of random coil conformations in water above pH 7.5 but at low pH adopts an antiparallel β-sheet conformation. It also self-assembles under physiological conditions in a concentration-dependent manner. Here we describe an unimer P(11)-4 production system and the use of a simple site-directed mutagenesis approach to generate a series of other P(11)-family peptide expression vectors. We have developed an efficient purification strategy for these peptide biomaterials using a simple procedure involving chemical cleavage with cyanogen bromide then repeated filtration, lyophilisation and wash steps. We report peptide-fusion protein yields of ca. 4.64 g/L and we believe the highest reported recovery of a recombinant self-assembling peptide at 203 mg/L of pure recombinant P(11)-4. This peptide forms a self-supporting hydrogel under physiological conditions with essentially identical physico-chemical properties to the chemically synthesised peptide. Critically it also displays excellent cytocompatibility when tested with primary human dermal fibroblasts. This study demonstrates that high levels of a series of recombinant self-assembling peptides can be purified using a simple process for applications as scaffolds in tissue engineering. |
format | Online Article Text |
id | pubmed-3111806 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Elsevier Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-31118062011-06-24 Recombinant self-assembling peptides as biomaterials for tissue engineering Kyle, Stuart Aggeli, Amalia Ingham, Eileen McPherson, Michael J. Biomaterials Article Synthetic nanostructures based on self-assembling systems that aim to mimic natural extracellular matrix are now being used as substrates in tissue engineering applications. Peptides are excellent starting materials for the self-assembly process as they can be readily synthesised both chemically and biologically. P(11)-4 is an 11 amino acid peptide that undergoes triggered self-assembly to form a self-supporting hydrogel. It exists as unimers of random coil conformations in water above pH 7.5 but at low pH adopts an antiparallel β-sheet conformation. It also self-assembles under physiological conditions in a concentration-dependent manner. Here we describe an unimer P(11)-4 production system and the use of a simple site-directed mutagenesis approach to generate a series of other P(11)-family peptide expression vectors. We have developed an efficient purification strategy for these peptide biomaterials using a simple procedure involving chemical cleavage with cyanogen bromide then repeated filtration, lyophilisation and wash steps. We report peptide-fusion protein yields of ca. 4.64 g/L and we believe the highest reported recovery of a recombinant self-assembling peptide at 203 mg/L of pure recombinant P(11)-4. This peptide forms a self-supporting hydrogel under physiological conditions with essentially identical physico-chemical properties to the chemically synthesised peptide. Critically it also displays excellent cytocompatibility when tested with primary human dermal fibroblasts. This study demonstrates that high levels of a series of recombinant self-assembling peptides can be purified using a simple process for applications as scaffolds in tissue engineering. Elsevier Science 2010-12 /pmc/articles/PMC3111806/ /pubmed/20932572 http://dx.doi.org/10.1016/j.biomaterials.2010.08.051 Text en © 2010 Elsevier Ltd. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license |
spellingShingle | Article Kyle, Stuart Aggeli, Amalia Ingham, Eileen McPherson, Michael J. Recombinant self-assembling peptides as biomaterials for tissue engineering |
title | Recombinant self-assembling peptides as biomaterials for tissue engineering |
title_full | Recombinant self-assembling peptides as biomaterials for tissue engineering |
title_fullStr | Recombinant self-assembling peptides as biomaterials for tissue engineering |
title_full_unstemmed | Recombinant self-assembling peptides as biomaterials for tissue engineering |
title_short | Recombinant self-assembling peptides as biomaterials for tissue engineering |
title_sort | recombinant self-assembling peptides as biomaterials for tissue engineering |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3111806/ https://www.ncbi.nlm.nih.gov/pubmed/20932572 http://dx.doi.org/10.1016/j.biomaterials.2010.08.051 |
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