Aluminium nitride - graphene dual-mode sensors for cancer cell detection
Lead Research Organisation:
Imperial College London
Department Name: Materials
Abstract
We propose to develop a technology for wafer scale device fabrication based on thin film heterostructures composed of graphene and aluminium nitride (AlN), which were recently demonstrated by our team for the first time. As a new orthogonal approach for biosensors, AlN-graphene heterostructures should enable simultaneous mass and charge detection of immobilized cellular and molecular species within one integrated device. This new capability is expected to lead to a dramatic reduction of false positives and false negatives in cancer cell detection based on antigen-antibody bond-mediated cell capturing. : As a bioactive sensor surface, bio-functionalized graphene enables charge detection via the electric field effect in graphene. Simultaneously, the graphene biointerface layer will be employed as an electrode - in combination with other graphene layer(s) embedded inside the heterostructure - to enable a fully integrated piezoelectric excitation and readout of mechanical deformations or oscillations of a free-standing AlN structure. Cell capture-induced alterations of the deformation or mechanical resonance should allow mass detection of the immobilized species. This orthogonal sensing capability of fully integrated devices will be tested and benchmarked for detection of cancer cells in blood or other body fluids.
Planned Impact
As discussed in detail within the Pathways to Impact statement, the research to be performed within the suggested project provides a new scientific and technological basis that has a realistic potential to revolutionize cancer diagnosis by body fluid analysis, and as such it will contribute to early state cancer diagnosis, cancer monitoring, therapy development and cancer research. We expect that during the course of this project we will be able to suggest pathways to novel diagnostic approaches, which will be subject to follow-up projects that include clinical trials. We intend to discuss potential applications on a regular basis with our industry stakeholders, Imperial Innovations and clinical partners. Moreover, proactive action will also be taken if results look promising in other application areas.
The suggested fit into Imperial's interdisciplinary research landscape and is in accordance with Imperial's Mission. Early cancer detection addresses one of the grand challenges of our modern society, which can only be tackled by a cross-disciplinary team like ours, which combines benchmarking research within Materials Science, Device Engineering, Theory and Simulation, Life Science and Cancer Research. As such, dissemination of any generated breakthrough by Imperial's internal media and external media connections is guaranteed. However, the main communication path of our results will be via peer reviewed publications in high-impact journals and conference contributions; prior to each, a careful check with respect to a possible patent application will be pursued in close partnership with Imperial Innovations and our industrial stakeholders.
The suggested fit into Imperial's interdisciplinary research landscape and is in accordance with Imperial's Mission. Early cancer detection addresses one of the grand challenges of our modern society, which can only be tackled by a cross-disciplinary team like ours, which combines benchmarking research within Materials Science, Device Engineering, Theory and Simulation, Life Science and Cancer Research. As such, dissemination of any generated breakthrough by Imperial's internal media and external media connections is guaranteed. However, the main communication path of our results will be via peer reviewed publications in high-impact journals and conference contributions; prior to each, a careful check with respect to a possible patent application will be pursued in close partnership with Imperial Innovations and our industrial stakeholders.
Organisations
- Imperial College London (Lead Research Organisation)
- University of Wroclaw (Collaboration)
- National Physical Laboratory (Collaboration)
- NanoView Biosciences (Collaboration)
- NanoFCM Co Ltd (Collaboration)
- Karlsruhe Institute of Technology (Collaboration)
- IMPERIAL COLLEGE LONDON (Collaboration)
- Link Microtek (United Kingdom) (Project Partner)
- EVA Diagnostics ltd (Project Partner)
Publications
Xu L
(2022)
On-chip integrated graphene aptasensor with portable readout for fast and label-free COVID-19 detection in virus transport medium.
in Sensors & diagnostics
Description | 1 . Development of technology for wafer scale manufacturing of graphene field effect transistor based biosensors: We have implemented and refined our in-house microfabrication technologie for wafer scale device fabrication, which includes patterning of of CVD graphene and optimization of contact resistance, as well as integration of microfluidics with graphene sensors. 2. Successful demonstration of various biomarker detection for a range of application using graphene field effect transistors: brain dementia biomarkers (results including first clinical results published), cancer exosomes and COVID virus like particles (publications in progress): As a viable alternative to immobilization of cells, we successfully demonstrated cancer exosome detection at concentration levels of medical relevance. Exosomes are cell vesicles which are released through the cell membrane onto the bloodstream and contain most of the genetic information of their mother cell (cell messengers). They are significantly smaller than cells (ca 100 nm diameter) , therefore exosome detection presents a viable alternative for early stage cancer detection from blood analysis. 3. Design and demonstration of a combined electric and acoustic biosensor based one a GFET (graphene field effect transistor) / SAW (surface acoustic wave) resonator device: As an important milestone of this project, we have developed and demonstrated a sensor based on a GFET with electrolyte gate electrode - combined with a surface acoustic wave microwave resonator made from a quartz substrate - with an intergrated microfluidic reservoir which contains the exosomes. We have demonstrated exosome detection with this dual mode device for the first time. 4. Enhanced antibody bonding by Carbon nanoparticles on top of graphene: carbon nanoparticles on the surtace of graphene were found to act as binding sites for antibodies, which lead to a much stronger bonding of target exosomes. According to initial findings this leads to a significant boost in detection sensitivity, which makes this method more attractive for early stage cancer detection by liquid biopsies. The results will be published in a high impact journal after further validation. 5, Realization of a highly selective cancer exosome detection using GFET biosensor arrays from a commercial foundry (Graphenea) 6. Demonstration of enhanced detection sensitivity using aptamer rather than antibody based immobilization. 7. Demonstration of GFETs for COVID-19 spike protein and antibody detection 8. Development of a compact electronic module for GFET multisensor analysis for point of care applications. 9. First clinical pilot study for pancreatic cancer in patient's blood plasma samples |
Exploitation Route | Our current work about GFET exposome detection is funded through a Primer Award by Cancer Research UK. As a results of this project, we have pursued a clinical study about cancer exosome detection in pancreatic cancer patients (publication under review). We have submitted a research proposal to the Alzheimer Society after first successful results about brain biomarker detection in blood plasma samples. |
Sectors | Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology,Security and Diplomacy |
URL | https://www.imperial.ac.uk/people/n.klein/research.html |
Description | A label-free and real-time Graphene bioSensor for exosome-driven point-of-care detection of early CANcers (Gr-SensorCAN) |
Amount | £97,224 (GBP) |
Funding ID | EDDCPT\100016 |
Organisation | Cancer Research UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 03/2021 |
End | 12/2021 |
Description | Electrodeposited 2D Transition Metal Dichalcogenides on graphene: a novel route towards scalable flexible electronics |
Amount | £434,446 (GBP) |
Funding ID | EP/V062387/1 |
Organisation | Engineering and Physical Sciences Research Council (EPSRC) |
Sector | Public |
Country | United Kingdom |
Start | 01/2022 |
End | 06/2024 |
Description | Advanced graphene device characterization |
Organisation | National Physical Laboratory |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | graphene deposition and device fabrication |
Collaborator Contribution | Kelvin probe microscopy, microwave near field microscopy |
Impact | mots recent joint publications: Adabi M, Lischner J, Hanham SM, Shaforost O, Wang R, Mihai, Hao L, Petrov P, Klein Nclose, Microwave study of field-effect devices based on graphene/aluminum nitride/graphene structures, Scientific Reports, ISSN: 2045-2322 (accepted) Gajewski K, Goniszewski S, Szumska A, Moczala M, Kunicki P, Gallop J, Klein N, Hao L, Gotszalk Tclose, 2016, Raman Spectroscopy and Kelvin Probe Force Microscopy characteristics of the CVD suspended graphene, DIAMOND AND RELATED MATERIALS, Vol: 64, Pages: 27-33, ISSN: 0925-9635 Goniszewski S, Adabi M, Shaforost O, Hanham SM, Hao L, Klein Nclose, 2016, Correlation of p-doping in CVD Graphene with Substrate Surface Charges, SCIENTIFIC REPORTS, Vol: 6, ISSN: 2045-2322 Gregory AP, Blackburn JF, Lees K, Clarke RN, Hodgetts TE, Hanham SM, Klein Nclose, 2016, Measurement of the permittivity and loss of high-loss a Near-Field Scanning Microwave Microscope, ULTRAMICROSCOPY, Vol: 161, Pages: 137-145, ISSN: 0304-3991 Goniszewski S, Gallop J, Adabi M, Gajewski K, Shaforost O, Klein N, Sierakowski A, Chen J, Chen Y, Gotszalk T, Hao Lclose, 2015, Self-supporting graphene films and their applications, IET CIRCUITS DEVICES & SYSTEMS, Vol: 9, Pages: 420-427, ISSN: 1751-858X |
Start Year | 2013 |
Description | Combined accoustic / electromagnetic biosensors, graphene biosensors |
Organisation | Imperial College London |
Department | Department of Chemical Engineering |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | - development of methodology and chip development for microwave detection of single cells - development of biofunctionalization of graphene layers - preparation of large area CVD layers for biuosensors |
Collaborator Contribution | - development of cell sorting by surface accoustic waves. - development of biofunctionalization of graphene layers |
Impact | - joint research proposal under preparation: combined accoustic / electromagnetic microfluidic device for marker free circulating tumoiur cell detection. - collaboration between postdocs and PhD students from both groups. - joint publication currently under reviews |
Start Year | 2016 |
Description | Exosome Characterization |
Organisation | NanoView Biosciences |
Country | United States |
Sector | Private |
PI Contribution | Providing exosome - on - graphene for characterization |
Collaborator Contribution | advanced optical imaging of exosomes |
Impact | - exosome characterization in progress |
Start Year | 2020 |
Description | Nanoparticles for enhanced graphene functionalization |
Organisation | Imperial College London |
Department | Department of Chemical Engineering |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We provided graphene samples |
Collaborator Contribution | They provided carbon nanopartices to strengthen the bonding of antibodies to the surface of graphene |
Impact | joint publication in progress |
Start Year | 2019 |
Description | Optical cancer exosome detection by flow cytometry |
Organisation | NanoFCM Co Ltd |
Country | China |
Sector | Private |
PI Contribution | Within the collaboration we have pursued cancer exosome detection by our graphene field effect transistor technology. These results were directly compared with flow cytometry measurements pursued by NanoFCM UK on the same samples. The results are discussed in a publication which is currently under review. |
Collaborator Contribution | Within the collaboration we have pursued cancer exosome detection by our graphene field effect transistor technology. These results were directly compared with flow cytometry measurements pursued by NanoFCM UK on the same samples. The results are discussed in a publication which is currently under review. |
Impact | publication submitted to ACS Nano currently under review |
Start Year | 2022 |
Description | Scanning transmission electron microscopy (STEM) on functionalized graphene |
Organisation | Karlsruhe Institute of Technology |
Country | Germany |
Sector | Academic/University |
PI Contribution | We provide samples of our graphene / functionalized graphene |
Collaborator Contribution | The group hold the record for high resolution of STEM. High resolution scanning transmission electron microscopy reveals the distribution of the self organized linker molecules on the graphene surface. |
Impact | STEM still in progress |
Start Year | 2018 |
Description | micromechanical graphene sensors |
Organisation | University of Wroclaw |
Country | Poland |
Sector | Academic/University |
PI Contribution | Free standing Graphene sensor structures, aluminium nitride thin films |
Collaborator Contribution | etched silicon microstructures for graphene sensor preparation, tunneling microscopy and surface analysis, piezoelectric measurements with atomic force microscopy. |
Impact | one joint publication: Gajewski K, Goniszewski S, Szumska A, Moczala M, Kunicki P, Gallop J, Klein N, Hao L, Gotszalk Tclose, 2016, Raman Spectroscopy and Kelvin Probe Force Microscopy characteristics of the CVD suspended graphene, DIAMOND AND RELATED MATERIALS, Vol: 64, Pages: 27-33, ISSN: 0925-9635 bilateral exchange of staff joint PhD supervision |
Start Year | 2014 |
Title | Clinical Pilot Study on PDAC (pancreatic ductal 34 adenocarcinoma) patients blood plasma samples using GFET sensor technology |
Description | Biosensors based on graphene field effect transistors (GFETs) have the potential to enable the development of point-of-care diagnostic tools for early-stage disease detection. However, issues with reproducibility and manufacturing yields of graphene sensors, but also with Debye screening and unwanted detection of non-specific species, have prevented the wider clinical use of graphene technology. Here, we demonstrate that our wafer scalable GFETs array platform enables meaningful clinical results. As a case study of high clinical relevance, we demonstrate an accurate and robust portable GFET array biosensor platform for the detection of pancreatic ductal adenocarcinoma (PDAC) in patients' plasma through specific exosomes (GPC-1 expression) within 45 minutes. In order to facilitate reproducible detection in blood plasma, we optimized the analytical performance of GFET biosensors via the application of an internal control channel and the development of an optimized test protocol. Based on samples from 18 PDAC patients and 8 healthy controls, the GFET biosensor arrays could accurately discriminate between the two groups, while being able to detect early cancer stages including grades 1 and 2. Furthermore, we confirmed the higher expression of GPC-1 and found that the concentration in PDAC plasma was on average more than one order of magnitude higher than in healthy samples. We found that these characteristics of GPC-1 cancerous exosomes are responsible for an increase in the number of target exosomes on the surface of graphene, leading to an improved signal response of the GFET biosensors. This GFET biosensor platform could facilitate the development of an accurate tool for the rapid diagnosis of early stage of pancreatic cancer. |
Type | Diagnostic Tool - Non-Imaging |
Current Stage Of Development | Initial development |
Year Development Stage Completed | 2022 |
Development Status | Actively seeking support |
Impact | Our biosensor platform based on GFET arrays has a strong potential for the detection of multiple biomarkers on one chip within a point of care setting. The detection of exosomes represent a promising new direction for early stage detection of cancer and other diseases. In our future research we are planning to refine the sensor technology for multi target detection, for example by employing machine learning for the readout of sensor arrays. We are considering larger clinical trials once the technology is fit for purpose. |