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Recombinant spider silk funcionalized with growth factors: investigation of retained funcionality
KTH, School of Biotechnology (BIO).
2016 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Recent advancements in studies on gene fusions of recombinant spider silk and a functional protein domain puts spider silk materials as a promising candidate in the research for more efficient delivery systems of growth factors (GFs) in the field of tissue engineering. This master thesis study investigates the functional properties of recombinant spider silk functionalized with growth factors. The retained functionalities of a gene fusion of the miniature decombinant silk protein 4RepCT and a fibroblast growth factor (FGF), as well as three different constructs of 4RepCT and epidermal growth factor (EGF), was investigated. Retained functionality of the silk domain was studied by investigating the formation of multilayers of the silk-FGF fusion protein using SPR and QCM-D technology, in addition to the ability of the gene fusions to form water stable fibers. Exposure of the FGF receptor (FGFR) binding site to FGF immobilized via silk was investigated by binding analysis of FGFR to a layer of 4RepCT-FGF fusion protein using SPR technology. Multilayer formation of the 4RecpCT-FGF fusion protein was demonstrated, as well as fiber formation of the different silk-GF fusions. The exposure of FGFR bindin sit to immobilized FGF via silk gave inconclusive results, as unspecific binding of FGFR to a layer of wild type 4RepCT was prominent. While the retained functionality of the sild domain was shown through multilayer formation and fiber formation, further studies need to be conducted on the retained funcionality of the growth factor domain.

Abstract [sv]

Fusionsproteiner bestående av rekombinant spindelsilke och en funktionell domän har i nyligen utförda studier visat sig kunna utgöra en lovande kandidat för mer effektiv leverans av tillväxtfaktorer inom vävnadstekniska applikationer. I detta examensarbete på avancerad nivå undersöktes egenskaperna hos rekombinant spindelsilke funktionaliserat med tillväxtfaktorer. Bibehållna funktioner hos fusionsproteiner bestående av ett miniatyr-silkeprotein (4RecpCT) och en fibroblast tillväxtfaktor (FGF), tillika tre olika fusioner av 4RepCT och epidermal tillväxtfaktor (EGF), studerades med SPR- och QCM-D-teknologi. Det bindande sätet hos en FGF receptor (FGFR) exponerades för FGF immobiliserat via silke för att undersöka inbindningen av FGFR till ett lager av 4RepCT-FGF. Uppbyggnad av multilager av lager av 4RepCT-FGF gav ofullständiga resultat, då ospecifik bindning till 4RepCT var framstående. Bibehållen funktion av silkes-domänen kunde visas genom multilager-bildning och fiber-bildning. Dock behövs ytterligare studier av bibehållen funktion av tillväxtfaktorn.

Place, publisher, year, edition, pages
2016.
Keyword [en]
spider silk, recombinants, growth factors, tissue engineering, biomaterial
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:kth:diva-180676OAI: oai:DiVA.org:kth-180676DiVA: diva2:895964
Supervisors
Examiners
Available from: 2016-01-20 Created: 2016-01-20 Last updated: 2016-01-20Bibliographically approved

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