Chemical protein engineering of elastin-like polypeptides (ELPs) as a potential bioadhesive scaffold
| dc.contributor.author | Poursan, Samane | |
| dc.contributor.examiningcommittee | Herbert, David (Chemistry) | |
| dc.contributor.examiningcommittee | Khajehpour, Mazdak (Chemistry) | |
| dc.contributor.examiningcommittee | Levin, David (Biosystems Engineering) | |
| dc.contributor.supervisor | Budisa, Nediljko | |
| dc.date.accessioned | 2024-01-08T16:26:08Z | |
| dc.date.available | 2024-01-08T16:26:08Z | |
| dc.date.issued | 2024-01-02 | |
| dc.date.submitted | 2024-01-03T21:32:57Z | en_US |
| dc.degree.discipline | Chemistry | en_US |
| dc.degree.level | Master of Science (M.Sc.) | |
| dc.description.abstract | Bioadhesives are essential for tissue adhesion and regeneration. Optimal bioadhesive should possess elasticity, robust adhesion capabilities in wet environments, and a cytocompatible scaffold for cell regeneration. Current options like cyanoacrylates and fibrin glues fail in these aspects due to cell toxicity and poor moisture adhesion, prompting the need for new materials that reconcile cytocompatibility with robust adhesion. These limitations by available commercial bioadhesives, demand the development of new adhesive biomaterials that combine cytocompatibility with strong adhesion. Marine organisms such as mussels secrete proteins that adhere effectively even underwater, but the adhesion mechanisms remain partly elusive. These proteins' adhesion strength is largely attributed to catechol side chains of L-Dopa, which can form various surface interactions, including hydrogen bonding and metal coordination. Commercial use of natural adhesive proteins is time- and labor-demanding, and uneconomical, as it takes thousands of mussel specimens to extract just one gram of adhesive proteins. In addition, low expression yield, low purification yield, and high levels of post-purification insolubility, restrict the production of recombinant foot protein types in Escherichia coli. To overcome these issues and inspired by intrinsically disordered domains of tropoelastin, the soluble precursor of elastin, elastin-like polypeptides (ELPs) consist of repeated sequences of the pentapeptide motif “VPGXG” (with “X” representing any amino acids except proline), emerge as promising synthetic alternatives. These stimuli-responsive adhesives exhibit a reversible phase transition regulated by temperature, making them suitable for applications such as drug delivery and wound healing. Their lower critical solution temperature (LCST) can be genetically adjusted to just below human body temperature, offering versatility for medical use. The bioadhesive efficacy of ELPs is linked to Dopa, crucial for adhesion, which is prone to unwanted oxidation during protein synthesis and purification. To counter this, we use a Dopa analog, meta-(ortho-(2-nitrobenzyl))-3,4-dihydroxyphenylalanine (m-oNB-Dopa), which is photocleavable and allows precise control over adhesion processes upon light irradiation. As a first step to achieving this goal, we designed two different constructs of ELP protein; one as a wild-type model protein containing tyrosine, valine, and lysine, as the “X” guest residues, and the other ELP construct containing amber stop codons replaced by tyrosine codons, was used to expand the scope of protein constructs. | |
| dc.description.note | February 2024 | |
| dc.identifier.uri | http://hdl.handle.net/1993/37950 | |
| dc.language.iso | eng | |
| dc.rights | open access | en_US |
| dc.subject | Bioadhesives | |
| dc.subject | Wound healing | |
| dc.subject | Elastin-like Polypeptides | |
| dc.subject | ELPs | |
| dc.title | Chemical protein engineering of elastin-like polypeptides (ELPs) as a potential bioadhesive scaffold | |
| dc.type | master thesis | en_US |
| local.subject.manitoba | no | |
| project.funder.identifier | NSERC: https://doi.org/10.13039/501100000038 | |
| project.funder.name | Natural Sciences and Engineering Research Council of Canada |