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Review article: Biosensors: Nanotools of Detection- A Review Payoshnee Bhalinge*, Sourab Kumar**, Abhishek Jadhav***,Shilpi Suman****, Pavan Gujjar*****, Nitesh Perla******
*Senior Lecturer, Dept of Oral & Maxillofacial Pathology and Microbiology M.A.Rangoonwala College of Dental Science, Pune – 411001 ** Senior Lecturer (Corresponding Author) Dept of Oral & Maxillofacial Pathology and Microbiology Dr.D.Y.Patil University, School of Dentistry, Sector 7, Nerul, Navi Mumbai, Maharashtra, India, ***Senior Lecturer , Dept of Oral & Maxillofacial Pathology and Microbiology , Dr.D.Y.Patil University, School of Dentistry, Sector 7, Nerul, Navi Mumbai, Maharashtra, India, ****PG Student , Dept of Oral & Maxillofacial Pathology and Microbiology , Dr.D.Y.Patil University, School of Dentistry, Sector 7, Nerul, Navi Mumbai, Maharashtra, India, *****Senior Lecturer , Dept of Oral & Maxillofacial Pathology and Microbiology , Narsinhbhai Patel Dental College and Hospital, Visnagar, Gujrat - 384315 *****Oral Pathologist and Consultant, Lifeline Charitable Trust, Sanpada, Navi Mumbai
Abstract: BIONSENSORS refer to ‘easy to use’ devices that are developed to help in the early diagnosis and treatment of disease. Early diagnosis is the key to successfully treating many diseases. Biosensors utilize the unique properties of biological and physical materials to recognize a target molecule and effect transduction of an electronic signal. The key advantages of biosensors are fast response and high sensitivity. Also, the basic advantage for point of care devices such as Biosensors would include integration of nano materials, microfluidics, automatic samplers, and transduction devices on a single chip. Biosensors are also being used as new analytical tools to study medicine. Thus this paper reviews how nano materials can be used to build Biosensors and how these Biosensors can help now and in the future to detect disease and monitor therapies. Nano technology will enable us to design sensors that are much smaller, less power hungry and more sensitive than current mico and macro sensors. (1,4) Keywords : Biosensor, diagnostic, detection, biochip
Introduction: organisms for foreign substances are unparalleled. Humans have been performing bio-analysis since Using bio-receptors from biological organisms or the dawn of time, using the sensory nerve cells of receptors that have been patterned after biological the nose to detect scents or the enzymatic reactions systems, scientists have developed a new means of in the tongue to taste food. As time has progressed, chemical analysis that often has the high selectivity so has our level of understanding about the function of biological recognition systems. These bio- of living organisms in detecting trace amounts of recognition elements in combination with various bio-chemicals in complex systems. Since biological transduction methods have helped to create the organisms are some of the most efficient machines rapidly expanding fields of bio-analysis and related ever created, scientists have sought to apply and technologies known as biosensors and biochips. (2) copy their efficiency for use in man- made creations. In particular, the recognition abilities of biological
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The emergence of nanotechnology is opening new urea in micro samples of undiluted whole blood or horizons for the development of nanosensors and plasma. Miniaturization also allowed additional nanoprobes with submicron-sized dimensions that analytical tools to be added to the biosensor, such as are suitable for intracellular measurements. (3) chromatography or capillary electrophoresis. The Although it is impossible to survey this entire newest generation of biosensors includes dynamic field, this issue presents articles about miniaturized multi-analyteimmunosensor devices some of the many types of biosensors and biosensor- with high-throughput capabilities and more than based applications to give the reader a sense of 1000 individually addressable electrodes per square importance and the enormous potential of these centimeter. These instruments can detect analytes devices. (3) present in the attomole range (3,8) Definition: Basic concepts of Biosensors Various definitions and terminologies are used As demonstrated in Fig. 1, a biosensor consists of a depending on the field of application. Biosensors are bio-element and a sensor-element. The bio-element also known as immunosensors, optrodes, chemical may be an enzyme, antibody, living cells, or tissue. canaries, resonant mirrors, glucometers, biochips The sensing element may be electric current, electric and biocomputers. (4) potential, and so on. (A detailed list of different Hence Biosensor can be defined as : possible bio-elements and sensor-elements is shown An analytical device which converts a biological in Fig. 2.) Different combinations of bio-elements response into an electrical signal (Biosensors, and sensor-elements constitute several types of Elsevier Applied Science). biosensors to suit a vast pool of applications. Or simply put : The bio elements and the sensor elements can be A device that uses specific biochemical reactions coupled together in one of the four possible ways mediated by isolated enzymes, immunosystems, demonstrated in Fig. 3, i.e. a) membrane tissues, organelles, or whole cells to detect chemical entrapment, b) physical adsorption, c) matrix / compounds usually by electrical, thermal or optical porous entrapment and d) covalent bonding. (4,9-12) signals. (3,5) a) In the membrane entrapment scheme, a History of Biosensors semi-permeable membrane separates the Father of the Biosensor analyte and the bio-element; the sensor is Dr. Leland C Clark (1918-2005) (6) (Refer to attached attached to the bio-element. table) - created many of the early biosensors in the b) The physical adsorption scheme is early 1960’s (3,7)using an ‘‘enzyme electrode’’ for dependent on a combination of van der Waals measuring glucose concentration with the enzyme forces, hydrophobic forces, hydrogen bonds, Glucose Oxidase (GOD). The success of single and ionic forces to attach the biomaterial to analyte sensor was followed by development of the surface of the sensor. integrated multi-analyte sensors capable of more c) The porous entrapment scheme is based on comprehensive analyses, such as a single instrument forming a porous encapsulation matrix for glucose, lactate, and potassium detection. around the biological material that helps in Technical developments in manufacturing enabled binding it to the sensor. the development of miniaturized integrated biosensors for determination of glucose, lactate, and
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International J. of Healthcare and Biomedical Research, Volume: 04, Issue: 03, April 2016, 26-39 d) In the case of the covalent bonding , the as an electrode (biological element).A thin sensor surface is treated as a reactive group layer of gold on a high refractive glass to which the biological materials can bind. surface can absorb laser light, producing The typically used bio-element enzyme is a large electron waves on the gold surface. Thus, protein molecule that acts as a catalyst in chemical binding of this target analyte to a receptor on reactions but remains unchanged at the end of the the gold surface produces a measurable reaction. Fig.4 shows the working principle of signal. enzymes. An enzyme, upon reaction with a Other Biosensors are based on laser light substrate, forms a complex molecule that, under launched into the fiber and the resulting appropriate conditions, forms the desirable product evanescent field at the tip of the fiber is used molecule releasing the enzyme at the end. The to excite target molecules bound to the enzymes are extremely specific in their action Fig.5. antibody molecules. A photometric detection This extremely specific action of the enzymes is the system is used to detect the optical signal basis of biosensors. (e.g., fluorescence) originating from the COMPONENTS of a Biosensor analyte molecules or from the analyte- There are 2 basic components of a Biosensor: (2,13) bioreceptor reaction.Such laser nanosensors 1. Biological Component can be used for invivo analysis of proteins a. Analyte (blood, serum, saliva, urine, and biomarkers in individual living cells.(3,14) stools, etc) 2. Electrochemical: Bionsensors based on b. Sensitive Bioelement created by biological enzymatic catalysis of a reaction that engineering(enzyme, antibody, nucliec produces or consumes electrons (the enzymes acid, cells, tissue, etc) used for the catalysis are called redox 2. Physical Component enzymes ). The sensor substrate contains a. Detector / Transducertransforms the three electrodes -Reference electrodes, signal resulting from the interaction of the working electrode, sink electrode. analyte with the biological element into The target analyte reacts with a working other signal that can be more easily electrode, and the ions produced create a measured and quantified potential which is referred to reference (Electrochemical, Piezoelectric, Optical, electrode to give a signal. Auxillary etc) electrode / counter electrode is a part of ion b. Amplifier / Display Unit is the signal source. processors or the electronic component The broad class of electrochemical sensors responsible for the display of the results encompasses both Amperometric devices,a in a user friendly way. high sensitivity biosensor that can detect Principles of detection electro-activespecies present in biological Biosensors work mainlyvia followingprinciples (2,3,13) test samples, and Potentiometric devices, 1. Photometric:Many of these Optical simple and sensitive and suitable for turbid Biosensorsare based on the phenomenon of samples without modification which can be surface plasmon resonance (SPR) which is an used for kinetic studies. (4,15) evanescent wave technique.Thisutilisesgold
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3. Piezoelectric: These acoustic 5. Thermal Detection Biosensors:exploit one devicesutilize crystals which undergo an of the fundamental properties of biological elastic deformation when an electric potential reactions, viz. adsorption or production of is applied to them. The alternating current heat that in turn changes the temperature of produces a wave in the crystal.The frequency the medium in which the reaction takes place. of wave is dependent on the elastic properties When the analyte comes in contact with the of the crystal. When this crystal is coated enzyme, the heat reaction of the enzyme is with biological element, this binding of the measured and calibrated against the analyte large target analyte to a receptor will produce concentration. The total heat produced or a change in frequency which will then give a adsorbed is proportional to the molar signal.The crystal used here is quartz. (13) enthalpy and the total number of molecules Mass sensing devices are also illustrated by in the reaction. The measurement of the quartz crystal microbalance. Target analytes temperature is typically accomplished via a for mass sensing can be either molecular thermistor and such devices are called components (such as proteins or nucleic enzyme thermistors.= acids) or whole cells.Mainly used for the Commonly used in the detection of pesticides detection of hybridized DNA, DNA-binding and pathogenic bacteria, its high sensitivity drugs and Glucose concentration.(15) to thermal changes without frequent 4. Ion Channel Switch: The use of ion recalibration and insensitivity to optical and channels has been shown to offer highly electrochemical properties in the sample are sensitive detection of target biological some of its advantages. (4) molecules. (16) By imbedding the ion channels Different types of biosensors in supported or tethered bi-layer membranes 1. Blood glucose biosensor (t-BLM) attached to a gold electrode, an } It is the biggest biosensor success story in electrical circuit is created. Capture commercial use for diabetes control; molecules such as antibodies can be bound to } Rapid analysis from a single drop of the ion channel so that the binding of the blood which is the analyte target molecule controls the ion flow through } Monitors glucose level in the blood (3,4) the channel. This results in a measurable 2. A Holographic Biosensor change in the electrical conduction which is } It uses holograms as sensing elements and proportional to the concentration of the is used to screen pancreatic disorder, by target. (13) detecting Trypsin in the duodenal fluid or Also called an enzyme field effect transistor a stool sample whichis possible by proper and primarily used for pH detection, it is use of bovine pancreatic trypsin inhibitor designed for Pathogenesis of Cancer (POC) (BPTI), an enzyme/bioelement; testing in critical care environments in } Potential applications in screening hospitals. Precise and quantitative test results pancreatic disorders at lower price; in an immediate timeframe, reduces the time } The major advantage is that very small of emergency diagnoses from hours down to trypsin levels can be detected within a 60- minutes.(4) minute period. (4)
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3. Nanoshell Biosensor 6. ImplantableBiosensor / In vivo } It uses gold as atransducer, because of its monitoring high scattering level, so that } To detect specific bio-molecules or subnanograms/mm quantities of Ig in chemicals in the body over a period of saline, serum, blood can be measured; time; } Itenhances chemical sensing by as much } Needles comprising of glucose electrode as 10 billion times and is being developed are placed subcutaneously. The receiver for applications including cancer receives the signal transmitted by the diagnosis, cancer therapy and testing of transmitter and displays the result.eg. proteins associated with Alzheimer’s glucose, urea, lactate, cholesterol.; disease. (3,18,19) } Micro-dialysis a most recent approach to 4. Biodetector an implantable biosensor works on the } It is used for DNA sequences and principle of mimicking the function of a mutation detection; blood vessel by implanting a micro- } The biodetectors are used to identify a dialysis probe into the tissue. small concentration of DNA (of } Nanowire biosensors can be decorated microorganisms like viruses or bacteria) with virtually any potential chemical or in a large sample; biological molecular recognition unit, } Biodetectors are many times faster than making the wires themselves analyte conventional PCR, more efficient in the independent. They are small, sensitive, number of DNA copies produced, label-free, real-time detection of a wide designed to use small volumes and range of chemical and biological species economical (3,4) helpful in array-based screening and in } Force Amplified Biological Sensor vivo diagnostics. (3,24,25) (FABS) also classified under bio- Application of Biosensors in Bio Sciences / detectors are extremely useful in the Medicine detection of anthrax, ricin, botulinum, and Biosensors are used as tools of early diagnosis in other pathogens. These can detect Bio Sciences / Medicine. Some of the applications various biologically active materials like are illustrated below: toxins, proteins, viruses, and bacteria in 1. Cancer Detection low concentrations. (4) The early diagnosis of cancer is the most 5. Immuno Biosensor critical factor for the survival of the patient } Used to detect Ag-Ab reactionfor analysis and theirtreatment. Rapid detection with an of complex fluids to analyze ultra-low detection limit of cancer markers is microorganisms, viruses, pesticides and crucial for the early diagnosis of cancer. industrial pollutants (20-23) Biosensors hence are instrumental in various } Designed for the ease of use by untrained ways in Cancer biology (1,3) personnel } Help to spot cancer cells circulating in the } Rapid assay and sensitivity comparable to blood either from a cancer patient, or that of ELISA (3)
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treated tumour that has returned again to 3. Other Uses cancerous phase; (26) Biosensors are used not only as diagnostic } Detects altered expression levels of and prognostic tools in medicine but have microRNAs (miRNAs), the new targets various other uses as given below: (13) for cancer therapy, that are correlated } Glucose monitoring in diabetes patients with caner type, tumour stage and which is a historical market driver; response treatments (46,47,48,49) } Pregnancy test which detects the hCG } Electrode Cell Impedance Sensors (ECIS) protein in urine; study the invasion properties of different } Routine analytical measurement of folic cancer cell types (1) acid, biotin, vitamin B12 and pantothenic } Nano-bio-chip (NBC) technique can be acid as an alternative to microbiological used for anylysis of oral cancer assay; biomarkers in exfoliative cytology } Protein microassays have been used to specimens targeting both biochemical and follow changes in Host (Humans) morphologic changes associated with proteins in response to a SARS infection early oral tumorigenesis. The alteration in (YIP, et al, 2005) while lectinmicroassays nuclear morphology and EGFR coupled to electrochemical detection have (epidermal growth factor receptor been used to distinguish different strains biomarker in oral squamous cell of E.Coli (Ertl, et al, 2003);(34) carcinoma) were successfully detected } Environmental applications e.g. the and quantified in NBC sensor, adequately detection of pesticides and river water reflecting cellular alterations in tumour contaminants; tissue even at early pre-malignant stages } Remote sensing of airborne bacteria e.g. making itcost effective and time efficient in counter-bioterrorist activities with detection time being seconds versus } Detection of pathogens;(35) hours/days with histology technique (27,28) } Determining levels of toxic substances 2. Biosensors in Orthopaedic Biology before and after bioremediation; The need for biosensors in orthopaedic areas } Detection and determining of is important (1,29-33) organophosphate; } for addressing health related problems } Force Amplified Biological Sensor associated with : (FABS) is Extremely useful in the o soft tissues including tendon and detection of anthrax, ricin, botulinum, and ligament, other pathogens and can detect various o hard tissue that is bone and biologically active materials like toxins, o bone-related diseases such as proteins, viruses, and bacteria in low osteoporosis and Paget’s disease concentrations. (4) } to estimate fracture risk and Biosensors with Saliva as an Analyte } to determine the response of bone to Saliva, an important medium to maintain health and treatment of bone disease disease, has a large number of diagnosticanalytes including steroid hormones (36) and HIV
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International J. of Healthcare and Biomedical Research, Volume: 04, Issue: 03, April 2016, 26-39 antibody (37) .Saliva is easier to handle for diagnostic development of biosensors utilizing new procedures because it does not clot, thus lessening technologies that offers improved sensitivity for the manipulations required. Salivary peroxidase detection with high specificity at the molecular levels are measured using an amperometric level, development of biosensor arrays that can electrode as an indicator of periodontal infections successfully detect, quantify and quickly identify (Ivnitski, et al, 2004) (15,38,39) . individual components of mixed gases and liquid in } Biosensors using saliva are helpful in an industrial environment. (3,15) detecting HIV-1, HIV-2, viral hepatitis It would be desirable to develop multiple integrated A,B,C oral cancer, breast cancer, type 2 biosensor systems that utilize doped oxides, diabetes, periodontal disease and polymers, enzymes or other components to give the caries;(42,43) system the required specificity. These should be low } The Oral Fluid Nano Sensor cost, disposable, reliable and easy to use as part of Test (OFNASET) is a handheld, an integrated sensor system. Sensing in picoliter to automated, easy-to-use integrated system attoliter volumes might create new problems in that will enable simultaneous and rapid development of micro reactors for sensing and novel detection of multiple salivary protein and phenomenon in very small channels (45) nucleic acid targets. OFNASET detector Nanotechnologies can provide label-free detection can be used in the office of a dentist or and are being applied to overcome some of the another health care provider for point-of- limitations of biochip technology. The real success care disease screening and detection.(44) depends on whether diagnostic techniques such as } Empowered with new and highly biosensors can be used to obtain reliable and sensitive technologies, the lower level of repeatable results in a timely manner.Bio-sensors analytes in saliva is no longer a may be wearable, implantable, provide real-time limitation. Almost anything that can be monitoring, communicate with drug injection measured in blood can also be measured instrumentation, and measure a drug release rate. (1) in saliva. (40,41) Conclusion Future issues in the development of Nano- As biosensor technology advances, the range of biosensor and Biochip applications broadens. Biosensors are now being New biosensors and biosensor arrays are being developed for detection of microbial pathogens and developed using new materials, nano-materials and their toxins, monitoring of glucose and other micro-fabricated materials including new methods metabolites, blood analysis, and other physiological of patterning. Biosensor components will use monitoring, cancer detection, and monitoring. As the nanofabrication technologies. Use of nanotubes, technology advances, producing lab-on-a-chip Buckminster fullerenes (buckyballs), silica and its devices, these self-contained portable instruments derivatives can produce nano-sized devices. Some of will allow measurements outside the laboratory, in the challenges in developing such biosensors will the field or at the bedside. Biochip technologies be,development of real-time non-invasive could offer a unique combination of performance technologies that can be applied to detection and capabilities and analytical features of merit not quantitation of biological fluids without the need for available in any other bio-analytical system multiple calibrations using clinical samples, currently available.(1,3)
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As the technology advances, producing microfluidic allow measurements outside the laboratory, in the devices aka lab-on-a-chip devices, which are field or at the bedsideis the next goal. inexpensive self-contained portable instruments that
Fig1
Fig2
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Fig3
Fig4
Fig5 Legend1: A schematic representation of biosensor Legend2: Elements of biosensor Legend3: In biomaterial-sensor coupling, the bio & sensor elements can be coupled together one of four ways : (a) membrane entrapment (b) physical adsorption (c) matrix entrapment and (d) covalent bonding Legend 4:working principle of enzymes Legend5: Specificity of enzymes is the basis of biosensor 3427 www.ijhbr.com ISSN: 2319 -7072 International J. of Healthcare and Biomedical Research, Volume: 04, Issue: 03, April 2016, 26-39
Table 1 History of biosensors YEAR HISTORY 1916 First report on the immobilisation of proteins; adsorption of invertase on activated charcoal 1922 First glass pH electrode 1956 Invention of the oxygen electrode (Clark) 1962 First description of a bio-sensor; an amperometric enzyme electrode for glucose (Clark) 1969 First potentiometric bio-sensor; urease immobilised on an ammonia electrode to detect urea 1970 Invention of the Ion-Selective Field-Effect Transistor (ISFET) (Bergveld) 1972/5 First commercial bio-sensor: Yellow Springs Instruments glucose bio-sensor 1975 First micro-based bio-sensor First immune-sensor: ovalbumin on a platinum wire
Invention of the pO 2 and cO 2optode 1976 First bedside artificial pancreas (Miles) 1980 First fibre optic pH sensor for invivo blood gasses (Peterson) 1982 First fibre optic based bio-sensor for glucose 1983 First Surface Plasmon Resonance (SPR) immuno-sensor 1984 First mediated amperometric bio-sensor: ferrocene used with glucose oxidase for the detection of glucose 1987 Launch of MediSenseExacTech TM blood glucose bio-sensor 1990 Launch of Pharmacia BIACore SPR – based bio-sensor system 1992 i-Stat launches hand-held blood analyser 1996 Glucocard launched Abbott acquires MediSense for $867 Million 1998 Launch of LifeScanFastTake blood glucose bio-sensor Merger of Roche and BoehringerMannhein to form Roche Diagnostics 2001 LifeScan purchases Inverness Medicals’ glucose testing business for $1.3 Billion 1999-current BioNMES, Quantum dots, Nanoparticles, Nanocantilever, Nanowire, Nanotube
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Table 2 Types of biosensor, and the biological materials used in their construction:-3,5,7 S.N. Type of Biosensor Type of Change Biological Example / Remarks Detected Component 1. Calorimetric / Temprature Enzyme Can detect temperature of Thermometric 0.0001C 2. Potentiometric Consumption of a Enzyme Urea detection gas 3. Amperometric Redoz reaction Enzyme Glucose biosensor Detect bacteria 4. Conductimetric Electrical Enzyme Urea detection conductivity 5. Optical Fluorescene / Enzyme, Ab, Luciferase for bacteria Absorbance DNA/RNA detection 6. Piezoelectric Resonant Antibodies Cocaine detection frequency 7. Whole Cell Based on 1,2,3 Live / Dead Nicotinic acid detection using biosensor microbial cells Lact. arabinosus
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