Journal
RSC ADVANCES
Volume 11, Issue 36, Pages 22334-22342Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/d1ra03584c
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Funding
- ARC Centre of Excellence for Nanoscale Biophotonics [CE140100003]
- MF and MH Joyner Scholarship in Science
- Norman and Patricia Polglase Supplementary Scholarship
- Research Training Program Stipend (RTPS)
- National Heart Foundation [102093]
- NHMRC Ideas grant [2001646, 2002254]
- ARC Linkage grant [LP 150100657]
- RMIT
- NHMRC Development grant [APP1178912]
- SA State Government
- National Health and Medical Research Council of Australia [2001646, 2002254] Funding Source: NHMRC
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A new coating procedure for optical fibre sensors suitable for protein analytes detection was presented in this study. The detection of streptavidin by binding to functionalised D-biotin was demonstrated, with stability to washing and long-term exposure to water. Different coating methods were employed to isolate SBP-biotin on the outer surface of the silk layer for optimal detection.
We 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 mu g 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.
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