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.
 
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 exosome detection by a graphene field effect transistor biosensor:
As a viable alternative to immobilization of cells, we successfully immobilized exosomes on graphene sensor surfaces and demonstrated specific exosome detection (publication currently under review) 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
Exploitation Route too early to say, but we are looking into different application of the technology which has been developed.
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 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 Graphenea Foundry for GFET biosensors 
Organisation Graphenea S.A.
Country Spain 
Sector Private 
PI Contribution collaboration and joint research
Collaborator Contribution Free GFET samples for test of biosensor
Impact ongoing collaboration about joint graphene biosensor development, joint publication 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 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