Synopsis presentation

1. Synopsis presentation PriyaSwethaDsouzaP Reg. No – 206/JAN 2016 JRF, YRC Yenepoya University

2. Title of the research Project Graphene based nanobiosensors Name and address of the student PriyaSwethaDsouza P Junior Research Fellow Yenepoya Research Centre Yenepoya University Name and address of the Research Guide Dr. Sudhakara Prasad Assistant Professor Yenepoya Research Centre Yenepoya University 21/06/2017

3. CONTENTS OF THE PRESENTATION Introduction Literature Survey Lacunae in the Literature Social Relevance Aim, Objectives and Hypothesis Methodology Study Plan Timeline Budget References

4. INTRODUCTION Early-stage diagnosis techniques play a vital role ‘Biosensor’ is one of the major advances in the field of healthcare science (Darwish et al., 2011) It can effectively serve as a low cost and highly efficient tool that uses biological entities like tissues, microorganisms, enzymes, antibodies, nucleic acids (NA) (DNA and RNA), cell receptors, synthetic ligandsetc (Mehrotra et al., 2016) Quickly, accurately, and reliably detect analyte molecules for medical diagnostics, environmental monitoring, defense, and food industry (Wang et al., 2016)

5. Graphene is a 2 dimensional layer consisting of sp2 hybridized • carbon atoms (Fuchs & Goerbig, 2008) • Graphene is the thinnest material ever known, impermeable to • gas, stronger than steel, has high surface area and is also • transparent (Huang et al., 2011) • One of the most promising applications for • grapheneinbiomedical technologies (Van, 2006) • Ideal material for single-biomolecule detection sensor platform • to detect specific target biomolecules (Yang et al., 2013)

6. Paper based diagnostic tools play a crucial role in the • development of simple point-of-care devices (Yetisen et al., • 2013) • The paper-based micro analytical devices (µ-PADS) are used • widely because of its advantages (Juntao et al., 2015) • Papers are used for fabricating µ-PADs • because the white colour serves as an • excellent background for the • colorimetric assays employing the paper as substrate (Liana et • al., 2012)

7. Literature Review • The journey of biosensors began in 1962, when Leland C. Clark Jr. and Cham Lyon demonstrated the first biosensor • Their concept of glucose enzyme electrodes enabled millions of diabetic patients to monitor their own blood sugar level • To prove their vision, they performed an experiment in which a thin layer of glucose oxidase(GOx) was entrapped at a Clark oxygen electrode by a semipermeable dialysis membrane • A decrease in oxygen concentration was found to be proportional to glucose concentration (Clark et al., 1962)

8. The advent of graphene in 2004, accelerated the biosensor research by adding new-dimensions in terms of high loading efficiency, good stability, biocompatibility, fast response time, low production cost, and consistent signal amplification even under the harsh environmental conditions that brings important advantages over many other nanomaterials (Geim et al., 2004) • A myriad of biological and chemical species such as proteins, viruses, bacteria, DNA, lipids, peptides, antibodies, metal ions etc. have been successfully detected by various graphene-based biosensingstrategies

9. Consequently, graphene continues to be a focus of research for the futuristic goal of multiplexed clinical diagnostic biosensors to provide early detection of many deadly diseases • Graphenederivative—its properties, production and assembly relevant to biosensingplatform • Structure • Large specific area • Electronic transportation • Ease of functionalization • Biocompatibility of graphene and its derivatives • Electrochemical behavior of graphene • Good adsorption capability

10. Schematic design representing the concept of ‘influence of surface-to-volume ratio’ on enhanced surface interactions between analyte molecules and different particle systems (microparticle, nanoparticle, and graphene) that eventually affects the overall electron conduction mechanism (Chauhan et al., 2017)

11. Schematic illustration of the sensing tactics involved in a graphene-based biosensor for in vitro and in vivo applications (Chauhan et al., 2017)

12. Schematic representation of (a) top-to-down, (b) bottom-to-up approaches involved in graphene synthesis. (c) Micromechanical cleavage. (d) Anodic bonding. (e) Photo-exfoliation. (f) Liquid phase exfoliation. (g) Growth on SiC.(h) Segregation/precipitation from carbon containing metal substrate. (i) Chemical vapor deposition. (j) Molecular beam epitaxy. (k) Chemical synthesis using benzene as building block (Chauhan et al., 2017)

13. Detection and sensing mechanisms • Almost every ‘nanoelectronic biosensor’ mechanism is based on the principle of charge-detection • When a biomoleculebinds to the sensor, the charge density of the sensor alters and an electrical sensing signal is generated • As graphene research progress rapidly, various types of binding receptors and ligands, physicochemical methods and nano-platforms have been used for the detection or control of different biological substances • This includes immune components, NA, specific proteins, DNA, biochemical compounds, glucose concentration, glutamine deficiency, hazardous fraction of metal ions and toxic gas

14. molecules present in the environment, detection of specific protein, aptamer, organophosphates and carbamic insecticides, species whole cells (like, cancer cells, stem cells, bacteria, or viruses), ascorbic acid (AA), uric acid (UA) level and dopamine (DA), hydrogen peroxide (H2O2), DNA and poly-lysine, pH, etc • These approaches have paved a new path in the development of biosensors having superior analytical performance, high sensitivity, high selectivity, LOD, low working potentials, and prolonged stability

15. LACUNAE IN THE LITERATURE Despite the developments, and reporting many sensors for various communicable diseases, a simple cost effective, non-invasive approach for the communicable disease detection is still lacking. In general, factors such as increasing prevalence of the pathogens, spread of disease, rising usage of home-based devices, and technological advancements are stimulating the demand for POC testing. Hence, the proposed simple, cost effective, non-invasive paper based analytical devices for detection of the communicable diseases; by utilizing the graphene oxide substrates are relevant in current scenario.

16. SOCIAL RELEVANCE Early diagnoses of diseases are very imperative in the timely treatment and prognosis. In addition, especially in the resource-limited settings there is a growing need for low-cost non-invasive methods to diagnose these diseases. So a low-cost sensing platform coupled with molecular biomarkers could be a solution for identifying diseases. The proposed research on the development of graphene based PADs are highly likely the need of the hour by considering the affordable public health care, and its easy disposable nature.

17. Rationale of the study Graphene oxide can quench photoluminescent compounds, based on their p-p interactions Known techniques are costly, require sophisticated instruments and require skilled person Simple, cost effective, disposable and user friendly methods are lacking

18. Aim of the study To develop graphene oxide based based paper analytical devices

19. Research Objectives • Fabrication of PADs, and surface functionalizationof GO/RGO with nano-bioconjugates for immunosensing. • Optical and surface morphological studies of hybrid graphene-nanobioconjugate composites, and immobilization onto PADs. • Bio-sensing applications by utilizing colorimetric and electrochemical techniques for infectious diseases, such as whooping cough, dengue fever, Listeriamonocytogenes and malaria. • Development of biosensor prototypes using artificial samples.

20. Methodology • Design and fabrication of µ-PADs Different approaches for creating two-dimensional μPADs. (a) Printing methods. (b) Mask-guided patterning methods. (c) Methods of creating physical boundaries. Abbreviations: LPAD, laminated paper-based analytical device; μPAD, microfluidicpaper-based analytical device; UVO, ultra-violet ozone treatment

21. Paper Substrate

22. Decoration of GO onto µ-PADs and standardization

23. Conjugation of Aptamers with GO and its surface characterization • The aptamers will be conjugated to the FAM dye and this 5' -dye modified aptamers are attached to graphene oxide through simple adsorption (p-p interactions)

24. Selective and sensitive detection of Listeriamonocytogenes using the fabricated µ -PADs • The detection of Listeriamonocytogenes is achieved by measuring the color change when the aptamer conjugated with chromogenic substrate and GO, interacts with the Listeria (imgaes taken by ZOE microscope and analyzed by ImageJ software

25. Expected Outcome Asimple low-cost approach for the early detection of Listeriamonocytogene The low-cost PAD device can revolutionize toxins detection in food industry and in the detection of communicable diseases The proposed sensing platform will have many application in other fields • This research will benefit the applicant, institution and society • This project will generate a foundation of knowledge on paper devices in India

26. Work plan

27. BUDGET ESTIMATES

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