Protein detection enabled using functionalised silk-binding peptides on a silk-coated optical fibre

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dc.contributor.authorCapon, P.K.
dc.contributor.authorHorsfall, A.J.
dc.contributor.authorLi, J.
dc.contributor.authorSchartner, E.P.
dc.contributor.authorKhalid, A.
dc.contributor.authorPurdey, M.S.
dc.contributor.authorMcLaughlin, R.A.
dc.contributor.authorAbell, A.D.
dc.date.issued2021
dc.description.abstractWe present a new coating procedure to prepare optical fibre sensors suitable for use with protein analytes. We demonstrate this through the detection of AlexaFluor-532 tagged streptavidin by its binding to D-biotin that is functionalised onto an optical fibre, via incorporation in a silk fibroin fibre coating. The D-biotin was covalently attached to a silk-binding peptide to provide SBP–biotin, which adheres the D-biotin to the silk-coated fibre tip. These optical fibre probes were prepared by two methods. The first involves dip-coating the fibre tip into a mixture of silk fibroin and SBP–biotin, which distributes the SBP–biotin throughout the silk coating (method A). The second method uses two steps, where the fibre is first dip-coated in silk only, then SBP–biotin added in a second dip-coating step. This isolates SBP–biotin to the outer surface of the silk layer (method B). A series of fluorescence measurements revealed that only the surface bound SBP–biotin detects streptavidin with a detection limit of 15 μg mL−1. The fibre coatings are stable to repeated washing and long-term exposure to water. Formation of silk coatings on fibres using commercial aqueous silk fibroin was found to be inhibited by a lithium concentration of 200 ppm, as determined by atomic absorption spectroscopy. This was reduced to less than 20 ppm by dialysis against water, and was found to successfully form a coating on optical fibres.
dc.description.statementofresponsibilityPatrick K. Capon, Aimee J. Horsfall, Jiawen Li, Erik P. Schartner, Asma Khalid, Malcolm S. Purdey ... et al.
dc.identifier.citationRSC Advances: an international journal to further the chemical sciences, 2021; 11(36):22334-22342
dc.identifier.doi10.1039/d1ra03584c
dc.identifier.issn2046-2069
dc.identifier.issn2046-2069
dc.identifier.orcidCapon, P.K. [0000-0002-7396-5757]
dc.identifier.orcidHorsfall, A.J. [0000-0003-1276-2742]
dc.identifier.orcidLi, J. [0000-0001-8818-6070]
dc.identifier.orcidSchartner, E.P. [0000-0003-1669-4302]
dc.identifier.orcidPurdey, M.S. [0000-0002-5063-8972]
dc.identifier.orcidMcLaughlin, R.A. [0000-0001-6947-5061]
dc.identifier.orcidAbell, A.D. [0000-0002-0604-2629]
dc.identifier.urihttp://hdl.handle.net/2440/131530
dc.language.isoen
dc.publisherRoyal Society of Chemistry
dc.relation.granthttp://purl.org/au-research/grants/arc/CE140100003
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/2001646
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/102093
dc.relation.granthttp://purl.org/au-research/grants/arc/LP150100657
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1178912
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/2002254
dc.rightsThis journal is © The Royal Society of Chemistry 2021.
dc.source.urihttps://doi.org/10.1039/d1ra03584c
dc.titleProtein detection enabled using functionalised silk-binding peptides on a silk-coated optical fibre
dc.typeJournal article
pubs.publication-statusPublished

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