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Label-free monitoring of proteins in optofluidic hollow-core photonic crystal fibres.
Heck, Jan R; Miele, Ermanno; Mouthaan, Ralf; Frosz, Michael H; Knowles, Tuomas P J; Euser, Tijmen G.
Afiliação
  • Heck JR; Department of Physics, Cavendish Laboratory, University of Cambridge, United Kingdom.
  • Miele E; Yusuf Hamied Department of Chemistry, University of Cambridge, United Kingdom.
  • Mouthaan R; Department of Physics, Cavendish Laboratory, University of Cambridge, United Kingdom.
  • Frosz MH; Yusuf Hamied Department of Chemistry, University of Cambridge, United Kingdom.
  • Knowles TPJ; Department of Physics, Cavendish Laboratory, University of Cambridge, United Kingdom.
  • Euser TG; Department of Electrical Engineering, University of Cambridge, United Kingdom.
Methods Appl Fluoresc ; 10(4)2022 Sep 20.
Article em En | MEDLINE | ID: mdl-36084629
The fluorescent detection of proteins without labels or stains, which affect their behaviour and require additional genetic or chemical preparation, has broad applications to biological research. However, standard approaches require large sample volumes or analyse only a small fraction of the sample. Here we use optofluidic hollow-core photonic crystal fibres to detect and quantify sub-microlitre volumes of unmodified bovine serum albumin (BSA) protein down to 100 nM concentrations. The optofluidic fibre's waveguiding properties are optimised for guidance at the (auto)fluorescence emission wavelength, enabling fluorescence collection from a 10 cm long excitation region, increasing sensitivity. The observed spectra agree with spectra taken from a conventional cuvette-based fluorimeter, corrected for the guidance properties of the fibre. The BSA fluorescence depended linearly on BSA concentration, while only a small hysteresis effect was observed, suggesting limited biofouling of the fibre sensor. Finally, we briefly discuss how this method could be used to study aggregation kinetics. With small sample volumes, the ability to use unlabelled proteins, and continuous flow, the method will be of interest to a broad range of protein-related research.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Soroalbumina Bovina / Fótons Idioma: En Revista: Methods Appl Fluoresc Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Reino Unido

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Soroalbumina Bovina / Fótons Idioma: En Revista: Methods Appl Fluoresc Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Reino Unido