Applications of Graphene Quantum Dots in Biomedical Sensors

Due to the proliferative cancer rates, cardiovascular diseases, neurodegenerative disorders, autoimmune diseases and a plethora of infections across the globe, it is essential to introduce strategies that can rapidly and specifically detect the ultralow concentrations of relevant biomarkers, pathogens, toxins and pharmaceuticals in biological matrices. Considering these pathophysiologies, various research works have become necessary to fabricate biosensors for their early diagnosis and treatment, using nanomaterials like quantum dots (QDs). These nanomaterials effectively ameliorate the sensor performance with respect to their reproducibility, selectivity as well as sensitivity. In particular, graphene quantum dots (GQDs), which are ideally graphene fragments of nanometer size, constitute discrete features such as acting as attractive fluorophores and excellent electro-catalysts owing to their photo-stability, water-solubility, biocompatibility, non-toxicity and lucrativeness that make them favorable candidates for a wide range of novel biomedical applications. Herein, we reviewed about 300 biomedical studies reported over the last five years which entail the state of art as well as some pioneering ideas with respect to the prominent role of GQDs, especially in the development of optical, electrochemical and photoelectrochemical biosensors. Additionally, we outline the ideal properties of GQDs, their eclectic methods of synthesis, and the general principle behind several biosensing techniques.DFG, 428780268, Biomimetische Rezeptoren auf NanoMIP-Basis zur Virenerkennung und -entfernung mittels integrierter Ansätz

Functionalization of gold and glass surfaces with magnetic nanoparticles using biomolecular interactions

Advances in nanotechnology have enabled the production and characterization of magnetic particles with nanometer-sized features that can be functionalized with biological recognition elements for clinical and biosensing applications. In the present study the synthesis and interactions between self assembled monolayers (SAMs) and functionalized nanoparticles have been characterized. Size and shape of magnetic nanoparticles synthesized wet chemically starting from ferrous and ferric salts were verified by transmission electron microscopy (TEM). These nanoparticles were then conjugated with FITC-labeled streptavidin through carbodiimide (EDC) chemistry. SAMs of thiol-capped biotins were synthesized on gold surfaces for capture of the conjugated nanoparticles. Characterization of nanoparticle functionalization and binding was performed using fluorescent microscopy, Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM) and scanning electron microscopy (SEM) with energy dispersive spectrometry (EDS). FT-IR spectra confirm the binding of biotin on gold via sulphur linkages. Fluorescent microscopy and XPS show streptavidin bound to the biotinylated gold surfaces. Elemental characterization from EDS indicates the binding of streptavidin-conjugated nanoparticles to biotinylated gold surfaces. Together, these techniques have application in studying the modification and behavior of functionalized nanoparticles for biological and other applications

Modification of Surfaces for Biological Applications

Understanding and controlling the nature of interactions at interfaces between various materials and systems has always been of interest, but with the fast development and need of new technologies it has become crucial to employ these interactions for various applications that range from biosensing of analytes in bodily fluids and the environment, to the development of bio-compatibatible and bio-mimicking surfaces that can be used to successfully couple biological systems to artificial materials and also build models for understanding biological systems better. Self-assembled monolayers (SAMs) are organized molecular assemblies that are formed by spontaneous adsorption of a compound in solution to a surface. They can change the surface properties without the need of changing the physical properties of the bulk material. Formation of SAMs on different substrates was investigated and performed in the work described in the thesis to be used in the detection of nucleic acids and enzyme inhibitors, development of surfaces with anti-adhesive and anti-microbial properties, development of surfaces for directed and patterned cell adhesion, and the construction of artificial membranes that can be used for studying the interaction of membrane proteins and the discovery of new pharmaceuticals. The surface of gold substrates was modified with alkanethiol compounds in order to attach biomolecules such as nucleic acids and proteins which allowed the modified surface to be used as a biosensor. Binding interactions were detected by electrochemical impedance spectroscopy and surface plasmon resonance. A surface resonance sensor provided a platform for the detection of DNA and RNA oligonucleotide sequences and also the detection of one-nucleotide mismatches from the hybridization these oligonucleotides. The same sensor platform, but with a different surface modification, was used to covalently attach an enzyme whose inhibitors are used as therapeutic drugs and also as pesticides and nerve agents. The sensor was able to detect two of these inhibitors, which are used in the treatments of Alzheimer\u27s disease, at a range of concentrations. This allowed the determination of binding affinity constants for the two inhibitors. The surface of gold was modified with functional groups in order to obtain inert surfaces with anti-adhesive properties with regard to the attachment of proteins. These surfaces are of interest in generating bio-compatible medical implants that can resist rejection from the host\u27s immune system andor the formation of bacterial biofilms. The inert property was combined with anti bacterial properties by attaching an antibiotic which is known to kill bacteria by binding to the cell membrane. Following characterization of gold surfaces by contact angle measurements, ellipsometry, grazing angle FT-IR, cyclic voltammetry and electrochemical impedance spectroscopy, the surface of glass substrates was modified with similar functional groups, by switching to a different coupling ligand for the substrate. Alkoxysilanes were used to modify the surface of glass, which can also be used to modify other materials, such as polymers and stainless steel. Gold and glass surfaces were also modified with antibodies, other proteins, and other functional groups which favored or prevented cell adhesion. This led to the ability for patterned and directed adhesion, and differentiation of several cell lines. Preparation and chemical modification of magnetic beads and the ability to modify the bead surface created the possibility to grow and trap cells in a flow-through magnetic bioreactor, which will be used for the continuous production of metabolites and growth of tissue in a three-dimensional construct. Modification of gold substrates also led to the construction of artificial phospholipid membranes, whose composition can be controlled and most importantly can be used for the insertion and characterization of membrane proteins on a two-dimensional platform. This will allow for characterization of ligand-protein and protein-protein interactions with surface characterization techniques such as surface plasmon resonance and electrochemical impedance spectroscopy. The various surface modifications and applications described in this work underscore a general theme that the surface of many different materials can be modified by using the correct functional groups for the formation of the self-assembled monolayer on the substrate surface, thus obtaining the same surface properties without the need to change the physical and chemical properties of the bulk material

Design and Development of Nanoconjugates for Nanotechnology

Nanotechnology builds devices from the bottom up with atomic accuracy. Among the basic nano-components to fabricate such devices, semiconductor nanoparticle quantum dots (QDs), metal nanocrystals, proteins, and nucleic acids have attracted most interests due to their potential in optical, biomedical, and electronic areas. The major objective of this research was to prepare nano-components in order to fabricate functional nano-scale devices. This research consisted of three projects. In the first two projects, we incorporated two desirable characteristics of QDs, which are their abilities to serve as donors in fluorescence energy transfer (FRET) and surface energy transfer (SET) as well as to do multiplexing, to engineer QD-based nanoconjugates for optical and biomedical applications. Immobilizing luminescent semiconductor CdSe/ZnS QDs to a solid platform for QD-based biosensors offers advantages over traditional solution-based assays. In the first project, we designed highly sensitive CdSe/ZnS QD SET-based probes using gold nanoparticles (AuNPs) as FRET acceptors on polystyrene (PS) microsphere surfaces. The emission of PS-QD was significantly quenched and restored when the AuNPs were attached to and then removed from the surface. The probes were sensitive enough to analyze signals from a single bead and for use in optical applications. The new PS-QD-AuNP SET platform opens possibilities to carry out both SET and FRET assays in microparticle-based platforms and in microarrays. In the second project, we applied the QD-encoded microspheres in FRET-based analysis for bio-applications. QDs and Alexa Fluor 660 (A660) fluorophores are used as donors and acceptors respectively via a hairpin single stranded DNA. FRET between QD and A660 on the surface of polystyrene microspheres resulted in quenching of QD luminescence and increased A660 emission. QD emission on polystyrene x microspheres was restored when the targeted complementary DNA hybridized the hairpin strand and displaced A660 away from QDs. The third project involved fabrication of different nanoconjugates via self-assembly of template-based metal nanowires and metal nanoparticles using oligonucleotides as linkers. These nanoconjugates can serve as building blocks in nano-electronic circuits. The template method restricted the oligonucleotides attachment to the tip of the nanowires. Nanowires tagged with hybridizable DNA could connect to complementary DNA-modified metal crystals in a position-specific manner

Nanostructured metallic film plasmonics: fabrication and biosensing applications

The research presented in this dissertation is interdisciplinary in nature. It covers the areas of micro- and nanofabrication, chemistry, materials science, and biological sensing. The running theme of the dissertation is the fabrication of micro- and nanostructures for use in plasmonic devices to aid in the optical detection of biomolecules. Phase I of the research focused on a bimetallic nanostructured (nanoslit) film to aid in improving the sensitivity in comparison to pure gold films. Phase II of the research investigated nanoledge structures (stair-step features) for their ability to trap biomolecules and aid in surface plasmon resonance sensing. Phase III of the research examined how to produce a fluidic dam, a microstructure with an overcut sidewall profile, which could aid in separating biological entities from the proteins of interest. Phase IV of the research assessed the use of the fluidic dam and nanoledge structures for detection of Troponin T, a biomarker used in the diagnosis of heart attacks. Phase V of the research focused on the design and microfabrication of a plasmonic device, which could study how surface plasmon resonance influences a photocurrent generated by immobilizing photosystem I in a nanoslit structure

Rapid Detection of SARS-CoV-2 Nucleocapsid Protein by a Label-Free Biosensor Based on Optical Fiber Cylindrical Micro-Resonator

The current global outbreak of coronavirus (COVID-19) continues to be a severe threat to human health. Rapid, low-cost, and accurate antigen detection methods are very important for disease diagnosis. The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) nucleocapsid protein (N-Protein) is often used as the diagnostic and screening for coronavirus detection. To this end, we propose and experimentally validate a highly sensitive whispering gallery mode (WGM) optical cylindrical micro-resonator (CMR) for bio immunoassay detection. To study the biokinetic process of immunoassay, the surface of the WGM micro-resonator is functionalized with N-Protein monoclonal antibody (N-Protein-m Ab), which led to the specific detection of N-Proteins. The spectral characteristics of the WGM resonance dip were investigated, and it is found that the transmission spectrum of WGM shows a monotonically increasing red-shift as a function of recording time. The WGM red-shift is due to the antibody-antigen reaction and can be used for the analysis of the immunoassay process. The wavelength shift is shown to be proportional to the concentration of N-Protein, which ranges between 0.1 and 100 μg /mL. Finally, different types of samples (concentration of 10 μg /mL of N-Protein) were prepared and tested to simulate the specificity of the sensor in the practical application environment. This method has the merits of a rapid assay, lower expense, easy preparation, and miniaturization, which makes the sensor have the potential for broad applications in the field of biochemistry and biomedical detection

Fabrication of Organosilane Nanostructures as Selective Sites for Surface Chemical Reactions

Naturally self-assembled mesospheres provide a practical route for controlling the arrangement of materials on surfaces at the nanoscale. Periodic arrays of well-defined nanostructures can be produced with different nanomaterials and interpattern spacings. Results presented in this dissertation demonstrate particle lithography methods developed for fabricating arrays of organosilane nanostructures. Surfaces were designed for the selective deposition of polymers and nanoparticles to produce multicomponent nanopatterns. The approaches for surface patterning provide new directions for studying surface chemistry at the molecular-level, and have practical application for emerging photovoltaic thin film technologies. Atomic force microscopy (AFM) provides unique capabilities for molecular visualization and ultrasensitive measurements of surface properties with nanoscale resolution. Organosilane nanopatterns bearing different functionalities and chain lengths were characterized using AFM to gain insight on molecular organization and surface-assembly processes. Indirect magnetic modulation (IMM) is a new instrument configuration for force modulation AFM that was developed for investigating mechanical properties of materials. The principle of IMM is based on indirect oscillation of soft nonmagnetic cantilevers through the tip holder assembly, which contains magnetic materials. Imaging can be performed in either ambient or liquid environments. The driving frequency for tip vibration can be selected to enhance contrast in amplitude and phase images, which provides information on the elastic response of thin-film materials. Images acquired with IMM furnish nanoscale resolution views of the morphology and elastic response of organosilane nanostructures. The dampening effect of liquid imaging media on cantilever oscillation during IMM was investigated using a liquid sample cell. Organic photovoltaic (OPV) devices are promising alternatives to traditional silicon based solar cells. A major challenge for OPVs is the requirement for higher efficiencies, or better device performance. The nanoscale morphology and molecular organization of the donor/acceptor materials in the organic layer affects the conductivity of OPV devices. To improve efficiency, new fabrication methods must be developed that are capable of controlling the molecular structure of the donor/acceptor materials. Using particle lithography combined with contact printing, billions of periodic and uniform pillar nanostructures of polythiophene can be fabricated on the surface. The dimensions and spacing can be selectively tuned by using different size latex masks

Emerging (Bio)Sensing Technology for Assessing and Monitoring Freshwater Contamination - Methods and Applications

Ecological Water Quality - Water Treatment and ReuseWater is life and its preservation is not only a moral obligation but also a legal requirement. By 2030, global demands will exceed more than 40 % the existing resources and more than a third of the world's population will have to deal with water shortages (European Environmental Agency [EEA], 2010). Climate change effects on water resources will not help. Efforts are being made throughout Europe towards a reduced and efficient water use and prevention of any further deterioration of the quality of water (Eurostat, European Comission [EC], 2010). The Water Framework Directive (EC, 2000) lays down provisions for monitoring, assessing and classifying water quality. Supporting this, the Drinking Water sets standards for 48 microbiological and chemical parameters that must be monitored and tested regularly (EC, 1998). The Bathing Water Directive also sets concentration limits for microbiological pollutants in inland and coastal bathing waters (EC, 2006), addressing risks from algae and cyanobacteria contamination and faecal contamination, requiring immediate action, including the provision of information to the public, to prevent exposure. With these directives, among others, the European Union [EU] expects to offer its citizens, by 2015, fresh and coastal waters of good quality