Periodic Reporting for period 4 - COSMOS (Control and measurement of single macromolecules in space and time)
Période du rapport: 2022-12-01 au 2024-05-31
We recently developed a new experimental approach to trapping a single molecule, such as a protein, in solution, based on its electrical charge. The proposal focuses on developing a novel measurement principle (escape time electrometry - ETe) for highly precise measurements of electrical charge on single molecules in the fluid phase. One of the main goals of the project is to use the ETe methodology to rapidly measure distributions in electrical charge of biomolecular species in solution. The potential application base is extremely broad with areas of immediate interest including detection of: (1) heterogeneous states of multimeric proteins, (2) antigen- Antibody interactions (3) small molecule binding to drug target proteins, (4) post-translational modifications such as phosphorylation, (5) conformational changes in biomolecules. The ETe measurement approach relies on optical observation of single molecules in a microscope and therefore requires the attachment of fluorescent labels to the molecule of interest. In a parallel sphere of activity we are also looking to develop the ability to develop an all electrical, label-free approach to the detection of the electrical properties of a single molecule in solution. The development of these new experimental approaches will not only enable researchers to address fundamental questions on the nature of molecular interactions and properties in solution but will also provide a new highly sensitive and precise biomedical detection technology that could have direct impact on aspects of societal health.
Conclusions of the action: The overall outcomes of the action have been extremely successful. We have invented a new biomolecular measurement technology, based on the originally proposed system, that achieves or is capable of achieving all the original goals envisioned. The fact that the project has attained a level of success and broad-spectrum relevance in a range of biomolecular analytics - corresponding arguably to one of the most optimistic scenarios at the outset - has been a truly remarkable outcome. Furthermore, in conducting further explorations of the basic science at the heart of the original application we uncovered the mechanism of a new, long ranged force at work between electrically charged objects in the solution phase. Yet again, the level of understanding achieved and insight attained during this phase have meant a truly unprecedented and unparalleled level of success. Both areas will now continue forward into future path breaking research streams, benefitting society both directly and indirectly through the generation of new knowledge and technological inventions with direct impact on health care and medicine.
Conclusions of the action: The overall outcomes of the action have been extremely successful. We have invented a new biomolecular measurement technology, based on the originally proposed system, that achieves or is capable of achieving all the original goals envisioned. The fact that the project has attained a level of success and broad-spectrum relevance in a range of biomolecular analytics - corresponding arguably to one of the most optimistic scenarios at the outset - has been a truly remarkable outcome. Furthermore, in conducting further explorations of the basic science at the heart of the original application we uncovered the mechanism of a new, long ranged force at work between electrically charged objects in the solution phase. Yet again, the level of understanding achieved and insight attained during this phase have meant a truly unprecedented and unparalleled level of success. Both areas will now continue forward into future path breaking research streams, benefitting society both directly and indirectly through the generation of new knowledge and technological inventions with direct impact on health care and medicine.
The implementation of the project in the first reporting period has been highly successful. My research group relocated from Zurich to Oxford as of 01.06.2018. The first few months of the project were devoted entirely to relocating and setting up the laboratory from scratch, re-building experimental set-ups and restarting experiments. One of the major fronts on which we had to invest significant effort was the transfer of our nanofabrication processes to the local cleanroom facilities.
This enterprise was successfully completed with the help of fantastic team work and significant amounts of time and effort on the part of the group members funded by the ERC. The recruited team started out with two doctoral (DPhil) students, and a postdoc working on theory and computation, all of whom transferred with the group from Zurich to Oxford. Within 6 to 9 months of moving to Oxford we had recruited two additional full-time postdocs and one part-time technician on the experimental side, and an additional PhD student with a focus on computation.
In the first phase of the project we were able to reproduce and improve upon previous electrometry measurements on single molecules in solution. In addition we counted a few major additional unpublished research achievements. We have: (1) demonstrated the ability to use electrometry to address questions related to molecular structure of nucleic acids like DNA and RNA, (2) developed a new experimental approach that facilitates the imaging of surface electrical charge distribution of material interfaces immersed in an electrolyte, and (3) made fundamental advances into the understanding key mechanisms underlying electrostatic interactions in solution. Overall, we laid the foundations for breakthroughs on three fronts comprising new technologies, fundamentally new molecular measurement ability and basic science discovery.
In the second half of the project, and towards the final phase, we had a major technological step forward that was not really anticipated prior to the project. We developed a way to use the same experimental platform use to measure charge in molecules to measure their size and shape with very high precision in the solution phase, even in samples with complex composition. We termed the method escape time size and shape spectrometry (ETsy). On a different front we carried out key experimental and theoretical work to demonstrate that electrostatic interactions in the fluid phase are far more elaborate and intricate than previously believed, thus opening doors to a major fundamental discovery in a key area of physics and chemistry, with immediate relevance in and impact on biology. The results from these two classes of breakthrough are in various stages of dissemination in the form of 5 additional papers that do not currently appear in the list of publications ensuing from the project. Furthermore the body of technological advances on the molecular measurement front are moving towards potential commercialisation. We are actively exploring avenues to bring the technology out to society potentially for use as an analytical tool in life sciences research and the pharma industry, but also exploring its potential in micro-chip based high-speed, high-sensitivity diagnostics.
This enterprise was successfully completed with the help of fantastic team work and significant amounts of time and effort on the part of the group members funded by the ERC. The recruited team started out with two doctoral (DPhil) students, and a postdoc working on theory and computation, all of whom transferred with the group from Zurich to Oxford. Within 6 to 9 months of moving to Oxford we had recruited two additional full-time postdocs and one part-time technician on the experimental side, and an additional PhD student with a focus on computation.
In the first phase of the project we were able to reproduce and improve upon previous electrometry measurements on single molecules in solution. In addition we counted a few major additional unpublished research achievements. We have: (1) demonstrated the ability to use electrometry to address questions related to molecular structure of nucleic acids like DNA and RNA, (2) developed a new experimental approach that facilitates the imaging of surface electrical charge distribution of material interfaces immersed in an electrolyte, and (3) made fundamental advances into the understanding key mechanisms underlying electrostatic interactions in solution. Overall, we laid the foundations for breakthroughs on three fronts comprising new technologies, fundamentally new molecular measurement ability and basic science discovery.
In the second half of the project, and towards the final phase, we had a major technological step forward that was not really anticipated prior to the project. We developed a way to use the same experimental platform use to measure charge in molecules to measure their size and shape with very high precision in the solution phase, even in samples with complex composition. We termed the method escape time size and shape spectrometry (ETsy). On a different front we carried out key experimental and theoretical work to demonstrate that electrostatic interactions in the fluid phase are far more elaborate and intricate than previously believed, thus opening doors to a major fundamental discovery in a key area of physics and chemistry, with immediate relevance in and impact on biology. The results from these two classes of breakthrough are in various stages of dissemination in the form of 5 additional papers that do not currently appear in the list of publications ensuing from the project. Furthermore the body of technological advances on the molecular measurement front are moving towards potential commercialisation. We are actively exploring avenues to bring the technology out to society potentially for use as an analytical tool in life sciences research and the pharma industry, but also exploring its potential in micro-chip based high-speed, high-sensitivity diagnostics.
By the end of the project we have developed new technologies that will enable for the first time rapid, high-throughput, high-precision measurement of a range of molecular properties in the fluid phase. The ETe/ETsy approach has developed into a mature experimental platform. In delivering high precision measurements of molecular charge, shape and size at high throughput and at the single molecule level, the developments constitute major progress in molecular measurement that are well beyond the state-of-the-art. We anticipate that the new opportunities for molecular measurement and detection that will emerge from the project will have significant impact both on fundamental research as well as on the biotechnology and pharmaceutical sectors. We further count an unanticipated and far-reaching fundamental discovery on the nature of intermolecular interactions in solution that has arisen from our efforts on the COSMOS project.