Sizing Up PD: Cavity-enhanced Characterisation of Single Biomarkers in Human Biofluids

Información del proyecto

SUPD

Identificador del acuerdo de subvención: 886216

DOI

10.3030/886216

Proyecto finalizado el 16 Abril 2024

Fecha de la firma de la CE 10 Marzo 2020

Fecha de inicio 2 Noviembre 2020

Fecha de finalización 23 Enero 2025

Financiado con arreglo a

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coste total

€ 289 732,80

Aportación de la UE

€ 289 732,80

Coordinado por

THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE
United Kingdom

Periodic Reporting for period 2 - SUPD (Sizing Up PD: Cavity-enhanced Characterisation of Single Biomarkers in Human Biofluids)

Período documentado: 2022-11-02 hasta 2023-11-01

Resumen del contexto y de los objetivos generales del proyecto

Measurements on single biomolecules provide a perspective of exceptional detail, enabling understanding of the underlying molecular mechanisms that govern biological processes. Most commonly, single-molecule measurements employ fluorescence which often requires the chemical attachment of fluorescent labels. As well as being perturbative to biomolecules and their interactions, labels limit the achievable resolution and sensitivity through complex photophysics. Recent developments in single-molecule label-free Rayleigh scattering based approaches have enabled the determination of the molecular weight of single biomolecules down to 30 kDa, however these techniques require proximity to surfaces. Microscale optical fiber based Fabry-Pérot cavities (FFPCs) provide a fully integrated route towards high sensitivity, label-free single-molecule measurements in the solution-phase. In this work we exploit both the small mode volume and high Q-factor of high reflectivity biconcave FFPCs to measure the properties of single proteins down to 1 kDa as they undergo Brownian motion in aqueous solution with signal-to-noise-ratios of >100. Here we improve the limit of detection 30-fold and the temporal resolution 100-fold compared to the current state-of-the-art. With this label-free technique we are able to determine the hydrodynamic radii of single proteins with unprecedented sensitivity. This technology has the potential to be developed into a highly sensitive diagnostic tool for complex diseases such as Alzheimer's and Parkinson's which require detection of small rare protein species. This has substantial implications for medical sciences and the ability to develop drugs and therapies for complex diseases.

Trabajo realizado desde el comienzo del proyecto hasta el final del período abarcado por el informe y los principales resultados hasta la fecha

Thus far in the project, we have developed a methodology for cavity-enhanced dynamic light scattering in optical microcavities. This has involved construction of an optical setup, fabrication of optical microcavities and performing single-molecule experiments with proteins. We have been able to measure signal from several protein species down to 1.2 kDa (0.75 nm), making it the most sensitive single-molecule technology in the world.

Avances que van más allá del estado de la técnica e impacto potencial esperado (incluida la repercusión socioeconómica y las implicaciones sociales más amplias del proyecto hasta la fecha)

This technology is currently >100-fold better than the current leading technology in single-molecule label free spectroscopy with vast potential for development and improvement. The small scale of the system will make it easily integrated into other setups. This has the potential to be a crucial technology in separation science, trace detection, and diagnostics.

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