An Optical Single Molecule Scanner of Protein Motion

Despite dramatic advances in x-ray crystallography and electron microscopy, we do not have a way to visualise functional proteins in motion.

This fellowship will lead the required breakthroughs and develop the first optical instrument to visualise proteins in real-time and at the level of single molecules. We propose to develop an instrument to probe single proteins in a specific and sensitive manner, while disturbing them as little as possible. The vision is to create a 'molecular scanner' that can characterise an arbitrary protein and its dynamics, a technology that is beyond the current state-of-the-art. Realising this sensor will lead to a new fundamental understanding of how the machinery of life functions.

The micro-optical sensor will allow us to analyse proteins in entirely new ways. We will be able to detect proteins specifically, from optically-induced vibrational motions, on portable coin-sized laboratories. The advances I envisage will result in a completely new approach for the analysis and diagnosis of protein-misfolding diseases (proteinopathies) such as prion diseases, Alzheimer's disease, Parkinson's disease, amyloidosis, and a wide range of other disorders. Our sensor platform will be able to contribute to the development of artificial molecular machinery by providing laboratory test beds that observe the motions of nano-machines in real time.

We will realise this instrument with optoplasmonic sensors. Optoplasmonic sensors enhance detection signals by reflection-driven circulation of the light. They concentrate the light at the nanoscale where they probe single proteins. We aim to scan the nanoscale light field across a single protein to provide information on the protein structure and its dynamics, resolving protein motions and vibrations at a temporal scale of nanoseconds and at a spatial scale of single bonds and atoms.

The optical technique developed in this fellowship will instigate entirely new domains in protein analysis. It will measure and visualise protein structure and its dynamics in-situ, in solution and at surfaces. It will accomplish one of the "holy-grails" of proteomics. Also, this technique can be integrated on a chip, allowing the identification of misfolded proteins from a trace amount of sample, with minimal sample preparation. Thereby it will create new analysis methods, biomarkers and standards for the pharmaceutical and chemical analysis industries.

A multitude of industries will be benefitted by the advances of this fellowship, including analytical sensing instrumentation, a $48.4 billion international market. The medical community desperately needs this analysis tool to rapidly detect and characterise intrinsically disordered proteins which cause the debilitating proteinopathies such as Parkinson's and Alzheimer's disease affecting more than 47 million worldwide, at an annual healthcare cost of ~$604 billion (WHO 2017).

Protein analysis is crucial to numerous scientific subfields of biology, chemistry, medicine, and pharmacology. A multitude of industries will benefit from the advances in protein sensing, including analytical sensing ($48.4 billion), cell analysis ($23 billion), chiral sensing ($9.2 billion), analysis instrumentation ($10.2 billion) and more (see below for details). This fellowship aims to revolutionize existing applications of protein sensing, instigating new domains with sensitivities beyond current limits. We anticipate high impact in several areas, especially protein analysis, analytical sensing, and protein structural studies.

Protein analysis:
Protein sensing is vital to a broad range of fields in medicine, biology, chemistry, and pharmacology. The analytical sensing instrumentation market is estimated to be e48.4 billion for 2016 (L1, and growing). We expect immediate impact in the fields of protein analysis, drug discovery, and clinical diagnostics with our emphasis on analysing proteins that cause Parkinson's and Alzheimer's diseases. In the long term, we expect a significant impact in the broader area of early diagnosis of dementia, which affects more than 47 million people worldwide, with 9.9 million new cases every year (WHO 2017, L2).
This fellowship will also lead to novel drug screening methods for the pharmaceutical industry (Roche etc.). The development of drugs that prevent proteins from misfolding can be a billion dollar international market. The analysis of disease-causing proteins on chip-scale devices will have considerable impact on the health industry, i.e. for the purposes of health monitoring and early diagnosis of proteinopathies. This can lead into rapid and cost-effective treatments of these diseases.

Analytical sensing:
The market of scientific instruments will be a $48.4 billion market by 2019 [L1]. This fellowship will introduce novel analytical methods and improve the limits of detection. Further developments of our technique can lead to a single cell protein analysis, which will significantly impact the cell analysis market. This market is expected to reach $23 billion by 2018 [L3].

Protein structural studies:
Understanding molecular structure and its dynamics is one of the holy grails in structural biology and medicine. Protein dynamics are extremely important for understanding and curing diseases (Parkinson's, Alzheimer's, familial amyloidosis and many more). Protein dynamics reveals the mechanisms of life's machinery and how it fails. The protein analysis instrumentation market that tackles these challenges was $10.2 billion in 2013 [L4]. The time-dependent, in-situ structural analysis of proteins will transform modern medical science. Current detection methods, such as NMR and CD/ORD do not have the detection limits required for time-dependent in-situ protein studies. Therefore, the proteins have to be extracted from their biological environment and cultivated in large quantities to facilitate the studies. The methods developed in this fellowship will allow for the first time the study of protein structure over time, without a label, and with minimal sample preparation. The extremely low detection limit enables the single-molecule and time-dependent detection of protein structure. This widens the potential of protein sensing in modern research. Also, especially in cases where sample material is scarce, the technology developed in this fellowship will enable the trace analysis of proteinaceous biomaterials. By constructing portable analysis devices, this fellowship will introduce protein sensing to various industry and government laboratories, and facilitate field studies that are planned with Roche.

L1 marketsandmarkets.com/Market-Reports/life-science-chemical-biotech-instrumentation-market-38.html
L2 who.int/mediacentre/factsheets/fs362/en/
L3 markets/andmarkets.com/Market-Reports/cell-analysis-market-157543946.html
L4 ...Market-Reports/proteomics-market-731.html