Shikha Sharma

Point-of-Care Diagnostics in Low Resource Settings: Present Status and Future Role of Microfluidics

The inability to diagnose numerous diseases rapidly is a significant cause of the disparity of deaths resulting from both communicable and non-communicable diseases in the developing world in comparison to the developed world. Existing diagnostic instrumentation usually requires sophisticated infrastructure, stable electrical power, expensive reagents, long assay times, and highly trained personnel which is not often available in limited resource settings. This review will critically survey and analyse the current lateral flow-based point-of-care (POC) technologies, which have made a major impact on diagnostic testing in developing countries over the last 50 years. The future of POC technologies including the applications of microfluidics, which allows miniaturisation and integration of complex functions that facilitate their usage in limited resource settings, is discussed The advantages offered by such systems, including low cost, ruggedness and the capacity to generate accurate and reliable results rapidly, are well suited to the clinical and social settings of the developing world.

Antibodies and antibody-derived analytical biosensors.

The rapid diagnosis of many diseases and timely initiation of appropriate treatment are critical determinants that promote optimal clinical outcomes and general public health. Biosensors are now being applied for rapid diagnostics due to their capacity for point-of-care use with minimum need for operator input. Antibody-based biosensors or immunosensors have revolutionized diagnostics for the detection of a plethora of analytes such as disease markers, food and environmental contaminants, biological warfare agents and illicit drugs. Antibodies are ideal biorecognition elements that provide sensors with high specificity and sensitivity. This review describes monoclonal and recombinant antibodies and different immobilization approaches crucial for antibody utilization in biosensors. Examples of applications of a variety of antibody-based sensor formats are also described.

Modulating electron transfer properties of gold nanoparticles for efficient biosensing.

Present study concerns modulating the electron transfer properties of gold nanoparticles through amino acid induced coupling among them. In addition to conductivity, the amino functionalization of the nanoparticles results in enhanced activity and operational stability of the biosensor fabricated using the same. Nanoparticles synthesized using amino acid as reducing agent (average diameter-20 nm), incorporate the natural coupling property of amino acids and are seen to align in a chain-like arrangement. The coupling of the individual nanoparticles to form chain like structure was confirmed by both absorption spectroscopy as well as transmission electron microscopy. The glucose biosensor developed by adsorption of glucose oxidase (GOx) enzyme onto these coupled gold nanoparticles showed enhanced efficiency as compared to the one with glucose oxidase immobilized onto gold nanoparticles synthesized using the conventional method (trisodium citrate as reducing agent). The fabricated biosensor demonstrated a wide linear concentration range from 1 μM-5mM and a high sensitivity of 47.2 μA mM(-1) cm(-2). Also, an enhanced selectivity to glucose was observed with negligible interference in the physiological range, from easily oxidizable biospecies, e.g. uric acid and ascorbic acid. Furthermore, the electrochemical biosensor has excellent long term stability- retaining greater than 85% of the biosensor activity up to 60 days.

Prostate cancer diagnostics: Clinical challenges and the ongoing need for disruptive and effective diagnostic tools

The increased incidence and the significant health burden associated with carcinoma of the prostate have led to substantial changes in its diagnosis over the past century. Despite technological advancements, the management of prostate cancer has become progressively more complex and controversial for both early and late-stage disease. The limitations and potential harms associated with the use of prostate-specific antigen (PSA) as a diagnostic marker have stimulated significant investigation of numerous novel biomarkers that demonstrate varying capacities to detect prostate cancer and can decrease unnecessary biopsies. However, only a few of these markers have been approved for specific clinical settings while the others have not been adequately validated for use. This review systematically and critically assesses ongoing issues and emerging challenges in the current state of prostate cancer diagnostic tools and the need for disruptive next generation tools based on analysis of combinations of these biomarkers to enhance predictive accuracy which will benefit clinical diagnostics and patient welfare.

Advances in ovarian cancer diagnosis: A journey from immunoassays to immunosensors.

This review focuses on the technological advancements, challenges and trends in immunoassay technologies for ovarian cancer diagnosis. Emphasis is placed on the principles of the technologies, their merits and limitations and on the evolution from laboratory-based methods to point-of-care devices. While the current market is predominantly associated with clinical immunoassay kits, over the last decade a major thrust in development of immunosensors is evident due to their potential in point-of-care devices. Technological advancements in immunosensors, extending from labeled to label-free detection, with and without mediators, for enhancing proficiencies and reliability have been dealt with in detail. Aspects of the utilisation of nanomaterials and immobilization strategies for enhancing sensitivity and altering the detection range have also been addressed. Finally, we have discussed some distinct characteristics and limitations associated with the recently commericalised technologies used for quantitation of relevant ovarian cancer markers.

Gold microwires based amperometric biosensor exploiting microbial architecture

Amalgamation of nanotechnology and biology has opened new horizons for controlled synthesis of nanomaterials of nano and micro-lengthscales for diverse sensing, catalytic and electromechanical applications. Inspired from nature and driven by the need to have nanostructures of desired morphology, microbial architecture has been exploited as a template in the present work. Biocompatible 1-D gold microwires, generated by assembly of amino acid functionalized AuNPs over the proliferating fungal hyphae, served as potential microelectrodes for electron transfer between enzyme and electrode surface. Delocalization of electrons over longer length scales, large surface area provided by assembled AuNPs and high biocompatibility yielded excellent analytical performance characteristics with high sensitivity of 43.2 µA/mM/cm(2) with standard deviation of 0.88% and wide linear range from 5 µM to 20 mM of glucose. The gold microwires thus generated demonstrate appreciable repeatability over 20 cycles in a cyclic voltammogram, and reproducibility with root mean square deviation as low as 1.3%. High stability and biocompatibility attribute these microwires with myriad potential biosensing and catalytic applications in varied domains.

ScFv-modified graphene-coated IDE-arrays for ‘label-free’ screening of cardiovascular disease biomarkers in physiological saline

Fatty-acid binding proteins (FABP) and myeloperoxidases (MPO) are associated with many chronic conditions in humans and considered to be important biomarkers for diagnosis of cardiac diseases. Here we assemble a new electrical biosensor platform based on graphene-coated interdigitated electrode arrays (IDE-arrays) towards ultrafast, label-free screening of heart type-FABP and MPO. Arrays of nanoscale (nanoIDE) and microscale (microIDE) electrode-arrays were fabricated on wafer-scale by combining nanoimprint and photolithography processes. Chemical vapor deposition grown multilayer graphene was transferred onto nano/microIDE-arrays and used as a high surface-to-volume ratio electrical transducer. Novel biofunctional layers of specially engineered anti-h-FABP and anti-MPO single-chain fragment variables (scFv) were immobilized onto graphene-coated IDE-array sensor platform for electrical detection of h-FABP and MPO in physiological saline. scFv fragments show increased sensitivity in comparison to the state-of-the-art competitive ELISA for their higher affinity towards target analytes. Deploying FABP and MPO specific scFvs as receptor molecules onto our high-sensitivity graphene-coated IDE-arrays with identical sensor characteristics and assays covering clinically relevant concentrations in physiological saline, we demonstrate realization of a simple and versatile biosensor platform capable of high performance cardiac-bioassays for point-of-care applications.

Immunoaffinity Chromatography: Concepts and Applications

Antibody-based separation methods, such as immunoaffinity chromatography (IAC), are powerful purification and isolation techniques. Antibodies isolated using these techniques have proven highly efficient in applications ranging from clinical diagnostics to environmental monitoring. Immunoaffinity chromatography is an efficient antibody separation method which exploits the binding efficiency of a ligand to an antibody. Essential to the successful design of any IAC platform is the optimization of critical experimental parameters such as (a) the biological affinity pair, (b) the matrix support, (c) the immobilization coupling chemistry, and (d) the effective elution conditions. These elements and the practicalities of their use are discussed in detail in this review. At the core of all IAC platforms is the high affinity interactions between antibodies and their related ligands; hence, this review entails a brief introduction to the generation of antibodies for use in immunoaffinity chromatography and also provides specific examples of their potential applications.

Strategies for overcoming challenges for decentralised diagnostics in resource-limited and catastrophe settings

ABSTRACT Introduction: Globally, both communicable and non-communicable diseases pose a serious threat to populations in developed as well as developing countries. Access to reliable diagnostic testing along with qualified health practitioners is severely limited in low resource and very remote areas and following natural catastrophes. Areas covered: This paper provides an overview of the challenges involved and suggests strategies to address them. The emergence of more robust, user-friendly, cost-effective and ‘sample-to-result’ point-of-care (POC) tools, along with the proliferation of mobile technologies, may provide a practical approach in addressing some of the challenges. Expert commentary: The successful implementation of POC testing requires the availability of versatile diagnostic technologies, improved platforms and back-up infrastructure, successful leveraging of human resources through training and, finally, engagement/coordination of associated stakeholders, including public health agencies, diagnostics companies, healthcare practitioners and local rural authorities.

Ultrasensitive direct impedimetric immunosensor for detection of serum HER2

Assesment of human epidermal growth factor receptor 2 status is a key factor prompting definitive treatment decisions that help in reducing mortality rates associated with breast cancer. In this article, highly sensitive and low-cost impedimetric immunosensor using single-chain fragment variable antibody fragments was developed for quantitative detection of human epidermal growth factor receptor 2 from serum employing gold nanoparticle-modified disposable screen-printed carbon electrodes. The gold nanoparticles facilitate fast electron transfer and offer a biocompatible surface for immobilization of small antibody fragments in an oriented manner, resulting in improved antigen binding efficiency. The single-chain fragment variable antibody fragment-modified screen printed immunosensor exhibits wide dynamic range of 0.01-100 ng mL-1 and detection limit of 0.01 ng mL-1. The advantages offered by this platform in terms of high sensitivity, broad dynamic range and low-cost demonstrates great potential for improved monitoring of human epidermal growth factor receptor 2 levels for the management of breast and other cancers.