Application of Nanobiosensors and Biochips in Health Care : A Review

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This review is an attempt to analyze the different applications of nanobiosenors and biochips in health care.
The biological and medical fields have seen great advances in biomolecules. This review is meant to provide an overview of the various types of biosensors and biochips that have been developed for biological and medical applications, along with significant advances over the last several years in these technologies. It also attempts to describe various classification schemes that can be used for categorizing the different biosensors and provide relevant examples of these classification schemes from recent literature.

1. Introduction
The emergence of nanotechnology is opening new horizons for the development of nanosensors and nanoprobes with submicron-sized dimensions that are suitable for intracellular measurements. A biosensor is defined as a µµdevice that uses specific biochemical reactions mediated by isolated enzymes, immunosystems, tissues, organelles or whole cells to detect chemical compounds usually by electrical, thermal or optical signals¶¶1. In recent years, many types of biosensors have been developed and used in a wide array of biomedical and other settings. Although it is impossible to survey this entire fast-moving field, this issue presents articles about some of the many types of biosensors and biosensor-based applications to give the reader a sense of the enormous importance and potential for these devices. One of the earliest references to the concept of a biosensor is from Dr. Leland C Clark who created many of the early biosensors in the early 1960¶s2 using an µµenzyme electrode¶¶ for measuring glucose concentration with the enzyme Glucose Oxidase (GOD). The success of single analyte sensors was followed by development of integrated multi-analyte sensors capable of more comprehensive analyses, such as a single instrument for glucose, lactate, and potassium detection. Technical developments in manufacturing enabled the development of miniaturized integrated biosensors for determination of glucose, lactate, and urea in micro samples of undiluted whole blood or plasma. Miniaturization also allowed additional analytical tools to be added to the biosensor, such as chromatography or capillary electrophoresis. The newest

and nanoscale biosensors. Modern fabrication techniques such as ink-jet printing. and the next type of devices to emerge may include miniature biosensors with high-density ligands. Biosensor that includes transducers based on integrated circuit microchips are often referred to as biochips. These instruments can detect analytes present in the attomole range3. Biosensors and biochips can be classified either by their bioreceptor or their transducer type ( Fig.1 Biosensing principle microcontact printing.1) illustrates the conceptual principle of biosensing process. and self-assembly continue to contribute to more advanced biosensors. selfcontained lab-on-a-chip capabilities. Nanobiosensors .2 Classification Schemes of Biosensors/Biochip 2. photolithography. (Fig. Fig. 2).generation of biosensors includes miniaturized multi-analyte immunosensor devices with highthroughput capabilities and more than 1000 individually addressable electrodes per square centimeter. Fig.

the SensiDx System reduces the time of emergency diagnoses from hours down to minutes. . realtime electrically based sensors for biological and chemical species4. Each chip contains multiple sensors. NY ) works by electronically detecting the binding of a target DNA molecule to sensors on a microchip.Nanomaterials are exquisitely sensitive chemical and biological sensors. Nanosensors with immobilized bioreceptor probes that are selective for target analyte molecules are called nanobiosensors. Biotin-modified SiNWs were used to detect streptavidin down to at least a picomolar concentration range. Their portability makes them ideal for pathogenesis of cancer (POC) applications but they can be used in the laboratory setting as well. This is the basis of the company¶s SensiDx System. By delivering precise and quantitative test results in an immediate timeframe. Nanowire biosensors Since their surface properties are easily modified. The target molecules form a bridge between two electrically separated wires. Australia). is based upon a synthetic self-assembling membrane. making the wires themselves analyte independent. a nanobiosensor device that has been designed for POC testing in critical care environments in hospitals. monitoring of metabolites in body fluids and detection of tissue pathology such as cancer. 2. In order to create a strong clear signal. which are metalized and can be seen by electron microscopy. 2. nanowires can be decorated with virtually any potential chemical or biological molecular recognition unit. Their applications include detection of microorganisms in various samples. These DNA wires µturn on¶ a sensor much like an on/off switch12. The nanomaterials transduce the chemical binding event on their surface into a change in conductance of the nanowire in an extremely sensitive. the bound target molecules are chemically developed to form conductive DNA wires.1. They can be integrated into other technologies such as lab-on-a-chip to facilitate molecular diagnostics. which can be observed by fluorescent imaging techniques. which greatly improves sensitivity. These chips now have billions of capture probes per sensor. The bridges. 2. The small size and capability of these semiconductor nanowires for sensitive. Electronic nanobiosensors The Biodetect system of Integrated Nanotechnologies ( Henrietta .3.2 Ion Channel Switch biosensor technologies The Ion Channel Switch (ICS). real-time detection of a wide range of chemical and biological species could be exploited in array-based screening and in vivo diagnostics. a novel biosensor technology of Ambri Ltd (Chatswood. which acts like a biological switch that detecting the signaling the presence of specific molecules by triggering an electrical current5. which can be independently addressed with capture probes for different target DNA molecules from the same or different organisms. Boron-doped silicon nanowires (SiNWs) have been used to create highly sensitive. are readily detected by measuring the resistance or other electrical properties of the sensor. A proprietary DNA Lithography process is used to attach capture probes to each of the sensors on the chip. real time and quantitative fashion. label-free.

6. PEBBLE can also be used for early detection of cancer. By using a magnetic field. PEBBLE nanosensors also show very good reversibility and stability to leaching and photobleaching.6.2 Laser nanosensors .1 Surface plasmon resonance technology Surface plasmon resonance (SPR) is an optical±electrical phenomenon involving the interaction of light with the electrons of a metal.6 Optical biosensors Many biosensors that are currently marketed rely on the optical properties of lasers to monitor and quantify interactions of biomolecules that occur on specially derived surfaces or biochips. These sensors are capable of real-time inter.2. Anti-HSV antibodies are conjugated directly to nanobeads using a bifunctional linker to avoid nonspecific interactions between medium components and protein G. as few as five viral particles can be detected in a 10 l serum sample. 2.5 PEBBLE nanosensors Probes Encapsulated by Biologically Localized Embedding (PEBBLE) nanosensors consist of sensor molecules entrapped in a chemically inert matrix by a microemulsion polymerization process that produces spherical sensors in the size range of 20 to 200 nm7.The nanobeads have a supramagnetic iron oxide core coated with dextran. little affected by light scattering and autofluorescence 8. In human plasma they demonstrate a robust oxygen sensing capability. The next generation microarray-based SPR systems are designed to help researchers profile and characterize biomolecular interactions in a parallel format. Surface plasmon resonance technology is the best-known example of this technology. Triangular silver nanoparticles have remarkable optical properties and their enhanced sensitivity to their nano environment has been used to develop a new class of optical sensors using localized SPR spectroscopy9. as well as very short response times and no perturbation by proteins. This system is more sensitive than ELISAbased methods and is an improvement over PCR-based detection because it is cheaper. 2. faster and has fewer artifacts. 2. Miniature optical sensors that specifically identify low concentrations of environmental and biological substances are in high demand.and intra-cellular imaging of ions and molecules and are insensitive to interference from proteins.4 Viral nanosensor Virus particles are essentially biological nanoparticles. Protein G is attached as a binding partner for antivirus antibodies. there is no optical sensor that provides identification of the aforementioned species without amplification techniques at naturally occurring concentrations. Currently. The optical±electronic basis of SPR is the transfer of the energy carried by photons of light to a group of electrons (a plasmon) at the surface of a metal. Herpes simplex virus (HSV) and adenovirus have been used to trigger the assembly of magnetic nanobeads as a nanosensor for clinically relevant viruses6 . 2.

7 Nanoshell biosensors Gold nanoshells have been used in a rapid immunoassay capable of detecting analyte within complex biological media without any sample preparation11. Nanoshells can enhance chemical sensing by as much as 10 billion times. serum and urine. When molecules and materials scatter light. A photometric detection system is used to detect the optical signal (e. chemists and other scientists to determine the nature of various materials. opening the door for new. There is also a demand to move clinical analysis from centralized laboratories to a doctor¶s clinic and patients self-testing at home.000 times more effective at Raman scattering than traditional methods. and the resulting evanescent field at the tip of the fiber is used to excite target molecules bound to the antibody molecules. TX ) have found that nanoshells are extremely effective at magnifying the Raman signature of molecules. That makes them about 10. etc. and whole blood. known as Raman scattering. This property. or for the detection of very small amounts of a material. An enormous limitation in the use of Raman scattering has been its extremely weak sensitivity. a property that directly leads to the enormous Raman enhancements observed. Nanoshells are already being developed for applications including cancer diagnosis. The metal cover of the nanoshell captures passing light and focuses it. fluorescence) originating from the analyte molecules or from the analyte-bioreceptor reaction10. cancer therapy. This tunability allows for the Raman enhancements to be optimized for specific wavelengths of light.In a laser nanosensor. a small fraction of the light interacts in such a way that it allows scientists to determine their detailed chemical makeup. Clinical applications of biosensors There has been a great demand for rapid and reliable methods which can be used in biochemical laboratories for determination of substances in biological fluids such as blood. Scientists at the Laboratory of Nanophotonics of Rice University ( Houston. reproducible enhancements of this effect. Successful detection was achieved in this system constitutes a simple immunoassay capable of detecting sub-nanogram-per-milliliter quantities of immunoglobulins in saline. and testing for proteins associated with Alzheimer¶s disease. and all-optical sensing applications. nanoshells can be tuned to interact with specific wavelengths of light by varying the thickness of their shells. like a few molecules of a deadly biological or chemical agent12. each individual nanoshell acting as an independent Raman enhancer. Aggregation of antibody/nanoshell conjugates with extinction spectra in the near infrared is monitored spectroscopically in the presence of analyte. something that could prove useful for engineers who are trying to probe the chemical processes within small structures such as individual cells. 3. That creates an opportunity to design alloptical nanoscale sensors±essentially new molecular level diagnostic instruments±that could detect as little as a few molecules of a target substance. Nanoshells can provide large. 2. Furthermore. drug designers.. clean. which could be anything from a drug molecule or a key disease protein to a deadly chemical agent. serum. is used by medical researchers.g. Most of the methods available in the market for rapid . Laser nanosensors can be used for invivo analysis of proteins and biomarkers in individual living cells. The finding that individual nanoshells can vastly enhance the Raman effect opens the door for biosensor designs that use a single nanoshell. laser light is launched into the fiber.

The glucometer is being adapted to the quantification of uric acid. triglycerides and phospholipids by using enzyme electrodes13. such as glucose. The instrument utilizes an enzyme electrode connected to a wick implanting to equilibrate with subcutaneous fluid. cholesterol and phospholipids. Similarly this ex vivo monitoring is used in sports medicine for measurement of lactate concentration.1. Therefore. Where there is a need of continuous monitoring of analytes. Recently. One of the approaches consists of an assembly of needles comprising a glucose electrode15 for subcutaneous use and the other approach is microdialysis. Recently. 3. Although biosensors have found immense applications in various fields.2. etc. such as lactate. their use in health care monitoring is of utmost importance. It was introduced in 1980 at the Centre of Scientific Equipment of the Academy of Sciences of the GDR. Ex-vivo monitoring A few instruments. dilutes it.detection are based on enzyme electrodes. saliva or skin has become popular. Uln) has been introduced as a commercial portable sensor for continuous glucose monitoring. which are in higher concentration in the extra cellular . The Glucometer GKM 01was the first commercial enzyme electrode based glucose analyzer developed in Europe . This instrument is helpful for obtaining longterm glucograms. Needle sensors have been implanted subcutaneously for several days and results are tele-transmitted to the receiver. Italy ) has been known for continuous measurement of glucose. measurement of metabolites in media other than blood has a great demand. Recently work on near infra red (NIR) method which is a reagent less system and non-invasive has started gaining interest. lactate and the activity of acetylcholine esterase. The substances. The probe essentially consists of a thin dialysis tube perfused with a sample (blood). therefore invasive biosensing sometimes proves to be very painful for the patients undergoing self-testing.14 have discussed the work carried out at labs in New Orleans and Rome for development of non-invasive sensors. glucose. Such types of measurements are care monitoring is of utmost importance. An amperometric urea sensor based on pH dependence of the anodic oxidation of hydrazine has been utilized in the glucometer GKM 02. the concept of non-invasive testing in sweat. An artificial pancreas µµbetalike¶¶ (EsaOte Biomedica. dialyze it and re-infuses blood cells into the blood stream and analysis of glucose concentration. In vivo monitoring Several approaches are known for in vivo measurements. The first enzyme electrode based lactate analyzer was developed in 1976 by LaRosch ( Switzerland) which used Cyt b2 on platinum electrode. It takes the blood from a patient vein. Lipid analyzer (ICA-LG 400) utilizes the enzyme electrode and is useful for determination of triglycerides. The transmitter converts current signal generated by the glucose needle biosensor to a very high frequency audio signal and the receiver demodulates back to a voltage. which are of great help in the treatment of continuous monitoring of diabetes and other metabolites. measurement of metabolites in media other than blood is important. namely cholesterol. etc. 3. Microdialysis is a more recent approach to an implantable biosensor and works on the principle of mimicking the function of a blood vessel by implanting a microdialysis probe into the tissue. pyruvate. They provide for a negligible enzyme consumption of <1 mg per sample. The lipid analyzer ICA-LG 400 from the Japanese company Toyo Jozo is capable of measuring whole group of analytes. urea. have been used for ex vivo monitoring. Guilbault et al. Another glucose sensor (Unitech. Geneoa .

Biosensors for health care 4. 4.17 reported that poly (3-dodecylthiophene) /stearic acid /glucose oxidase (P3DT/SA/GOX) Langmuir±Blodgett films based glucose biosensor can be used for at least 35 measurements and was found to be stable upto 40 days. The level of the glucose can be monitored either in vivo or in vitro. These electrodes have been shown to be useful for glucose estimation from 1 to 40 mM and stability of about 6 days. Most of the urea biosensors available in literature are based on detection of NH4+ or .1. heart failure. The product is available with the Indian markets for the consumers. 18. Ramanathan et al. Two different technologies have been approached for the development of miniaturized systems. disposable type sensors were developed for the purpose of on-line analysis25. 4. which can be used as either implantable catheter type devices or for in vivo monitoring in combination with microdialysis system23. Recently. 26. glucose. potassium. The i-STAT portable clinical analyser which is a significant commercially available biosensor. Other approaches. India has recently developed a screen printed electrode based lactate biosensor. can measure a range of parameters: sodium. which have been proposed for in vivo monitoring include enzyme based electrochemical. electrochemical and optical approach. Group at the National Physical Laboratory. The sensor was found to have a diminished enzyme activity (about 10%) and leaching of the enzyme from the matrix. The first approach for in vitro study was pioneered by Shichiri et al. Urea and creatinine biosensors Urea estimation is of utmost importance in monitoring kidney functions and disorders associated with it. Thin film electrodes have been developed. blood urea nitrogen (BUN) and haematocrit. Many biosensors have been reported to date21.3. Such a disposable lactate sensor has a linear dynamic range from 0. chloride. Singhal et al.2 to 1 mM of lactate and stability of about 3weeks. enzyme based field effect transistor (ENFET).19 covalently attached glucose oxidase on poly (o-amino benzoic acid) and fabricated the screen printed electrodes made of this material. shocks. 24. 22. Glucose biosensor Detection of glucose has been the most studied analyte in diabetic patients. India is based on the screen printed graphite electrode having a mediator incorporated in the working electrode. the concentration can be determined by coupling it with a biosensor.fluid outside the probe. The sensors are fabricated using thin film microfabrication technology on a disposable cartridge20.15Mascini et al.16 reported an µµartificial pancreas¶¶ for continuous measurement of glucose. Li and coworkers28have recently reported the sol±gel encapsulation of lactate dehydrogenase for optical sensing of L-lactate.2 Lactate biosensor Lactate measurement is helpful in respiratory insufficiencies. The NPL Glucosense developed at the National Physical Laboratory. 4. defuse in as soon as the substances are carried out of the body by the perfusion liquid. enzyme based thermoelectric. 19 . metabolic disorder and monitoring the physical condition of athletes. A number of glucose biosensors have been reported which are based on conducting polymers 17. Secondly.

or immobilization involving avidin± biotin complexation51 were adopted for a DNA probe to the surface of an electrochemical transducer. pyruvic acid and uric acid. L-ascorbic acid. Because of its simplicity. gold53-55 .28 have recently immobilized urease on poly (Nvinyl carbazole/stearic acid). 4. Uric acid biosensor Uric acid is one of the major products of purine breakdown in humans and therefore its determination serves as a market for the detection of range of diosorders associated with altered purine metabolism. Various attempts have been made to develop a biosensor for the estimation of uric acid41-47. The sensor was found to be responsive even in the presence of potential electrical interferents. Osaka et al. or conducting polymer56. hyperthyroidism. Conventional methods for the analysis of specific gene sequences are based on either direct sequencing or DNA hybridization48. hyperuricaemia and Lesch±Nyhan Syndrome.4.59 linked the tagged DNA to the surface of the microsphere using a suitable reagent. ischemia. anemia and coronary artery diseases. Vengatajalabathy and Mizutani40 demonstrated an amperometric biosensor for cholesterol determination by a layer-by-layer selfassembly using ChOx and poly (styrenesulfonate) on a monolayer of microperoxidase covalently immobilised on Au-alkanethiolate electrodes.33 presented a cholesterol biosensor by co immobilization of cholesterol oxidase and peroxidase on sol±gel films and utilized these films for estimations of cholesterol. kidney injury. 57. most of the traditional techniques in molecular biology are based on hybridization. etc. such as horseradish peroxidase58.31 constructed a highly sensitive and rapidflow injection system for urea analysis with a composite film of electropolymerized inactive polypyrrole and a poly ion complex. hypertension. notably gout. Another effort is the use of microfabrication system and micro . 30. 4. Gambhir et al. Kumar et al. as a reducing agent uric acid scavenges free oxygen radicals. covalent attachment50. In the case of a common sandwich assay the signal generating species is an enzyme. Reports on the development of cholesterol biosensors are available33-39. Recently. Determination based on the inherent specificity of an enzymatic reaction provides the most accurate means for obtaining true blood cholesterol concentration. Elevated levels of uric acid are observed in a wide range of conditions such as leukaemia. Additionally. Cholesterol biosensor Determination of cholesterol is clinically very important because abnormal concentrations of cholesterol are related with hypertension .HCO3¯sensitive electrodes28. Rapid detection of pathogenic infections and screening of cDNA colonies are required in molecular biology. pneumonia. preventing their destructive action towards tissue and cells. Singhal et al.5. Several immobilization techniques such as adsorption49. 4. 29.32 have recently co-immobilized urease and glutamate dehydrogenase on electrochemically prepared polypyrrole/polyvinyl sulphonate for the fabrication of urea biosensor. Lund et al. DNA biosensor DNA biosensors have an enormous application in clinical diagnostics for inherited diseases. The transducer was made from carbon52.6.

Biosensor Transducers The transducer converts the biochemical interactions into a measurable electronic signal.2 Indirect detection biosensors The second class of transducers. carbazole as a hapten. 5. They believed that anion doped polypyrrole undergoes ion exchange with PO4¯ of DNA to facilitating the adsorption. The most common direct detection biosensor systems employ evanescent wave. which may react to modulate the polymer electrochemistry.g. Dr. In the recent past. technology which measures resonant oscillation of electrons on the surface of a metal. pesticides and industrial pollutants have been developed64-67. Barnett et al. have detected thaumatin using antibody containing polypyrrole electrodes67. a number of methods for immunoassay by specific interactions between antibodies and antigens to analyze microorganisms.mechanical technology to the preparation of DNA samples and their analysis (e.1. typically use non-catalytic ligands such as cell receptors or antibodies. 5. electrooptical. Presently. Recently. viruses. Immuno-sensors are the analytical systems based on immuno-chemical principles that can automatically carry out estimation of desired parameter. DNA probes and biosensors have widely attracted attention for diagnosis of various disorders60-63. antibodies have been raised against the conducting polymer. DNA chip). indirect detection sensors. 4. 5. The transducer works either directly or indirectly. During the past decade. Electrochemical. in which the biological interaction is directly measured in real time. relies on secondary elements that are often catalytic elements such as enzymes. rapid assay and sensitivity comparable to that of ELISA.7. portable instruments for analysis of complex fluids and are designed for the ease of use by un-trained personnel. or surface plasmon resonance (SPR). acoustical. Vincent Gau¶s and Dr. Immuno-sensors Immuno-sensors are small. Omowunmi Sadik¶s groups describe the use of electrochemical transducers to measure the oxidation or reduction of an electroactive compound .57 have recently attempted to immobilize DNA on conducting polypyrrole/ polyvinyl sulphonate films and demonstrated the adsorption characteristics. and mechanical transducers are among the many types found in biosensors. Some examples of secondary elements are the enzyme alkaline phosphatase and fluorescently tagged antibodies that enhance detection of a sandwich complex. Direct detection biosensors Direct detection sensors. immunoassays have relied on complex indirect enzyme methods in which the resultant product of the enzyme immuno reaction can be measured. Gambhir et al. It has been observed by cyclic voltammetry that the reaction of the antiserum influences the polymer matrix electrochemistry by an amperometric response.

Frances Ligler¶s group has used optical fluorescence to develop a multi-analyte indirect detection biosensor. DNA amplification. there is significant research on manufacturing to produce advanced integrated devices. µµLab-on-a-chip¶¶ biosensors contain integrated microfabricated fluidics systems and are designed to perform multi-step high-resolution biological or chemical assays. flow cytometry and complex biochemical reactions. or glass chip. is also used for indirect detection2. the amount of reagents. and costs. microfluidic circuits can be designed to perform many biochemical processes including immunological assays. Fluorescence resonance energy transfer (FRET). Table 1:Various biosensor transducers. light-addressable potentiometric sensors (LAPS) combine electrooptics and electrochemistry for indirect detection. measuring fluorescence of the secondary ligand. As discussed in the article by Dr. Other common indirect detection biosensors employ optical fluorescence. labor. organelle. These devices can contain many channels. quartz. Dr. a process where energy from an excited fluorophore is transferred to a neighboring acceptor molecule. allowing for massively parallel biochemical processes and multi-analyte detection3. Tatsuo Yoshinobu¶s group. as discussed by Dr. manipulation. 5. principles and applications86 Transducer system Principle Applications Enzyme electrode Amperometric Enzyme substrate and immunological system Conductometer Conductance Enzyme substrate Piezoelectric crystal Mass change Volatile gases and vapors Thermistor Calorimetric Enzyme. Steven Soper¶s and Fred Battrell¶s groups.on the secondary ligand in one common type of indirect detection sensor. Finally. Many of these devices are fabricated using molding or photolithographic processes developed in the microelectronics industry to create circuits of chambers and channels using composite materials. Integrated biosensors and lab-on-a-chip Along with the development of better ligands and alternative transducer technologies. Their small dimensions and large relative surface area within the fluidic system reduce reaction times. silica. whole cell or tissue sensors for . and analysis.3.

pollutants. enzyme substrates and immunological analytes 6. Since the most limiting factor is the ratio between the number of data points per day and the cost per data point. enzyme electrodes. which is functionalized with an array of probes (features). this first mass-selection is best done at a molecular level where DNA-chips and protein microarrays find their most common application. After that the investigations will move to the whole cells. bring the analytes in contact with the probes. vitamins. i.substrates. The first two parameters reflect the ability to deliver a correct response at different . the immobilization of different DNA sequences onto different positions (probes). 6. and offer many advantages over the conventional analytical methods. These advantages become very evident if we consider the workflow during a typical drug screening process. antibiotics. by flooding the chip with the sample solution and let the hybridization take place. Ion sensitive electrode (ISE) Potentiometric Ions in biological media. gases. wide dynamic range and high specificity. A single probe contains identical molecules. the array is scanned to detect on which probes a hybridization took place. Depending on the different fields of application. 3) Readout.1 DNA Biochip A DNA-chip consists in most cases in a glass or quartz slide acting as a carrier.e. the requirements for the DNA-chip may include high sensitivity. enzyme substrates and immunological systems Optoelectronic/wave guide and fiber optic device Optical pH. where the Channelomics can deliver very clear informations about the effect of drugs on the physiology of the cells68. gene expression for drugs screening. or medical diagnosis of cancer or genetic diseases. Products. etc. after washing the chip to remove all non-bound molecules. ii) the required sample quantities are minimal. for this reason it is also called feature.g. gases. Biochip Biochips are the basis for miniaturized biochemical assays. 2) Hybridization. e. the most significant of which are: i) a variety of analytes can be investigated simultaneously in the same sample. iii) low consumption of scarce reagents. At the beginning there is the need for a selection of few eligible compounds out of a large variety of molecules for a given purpose. enzyme immunosensors Field effect transistor (FET) Potentiometric Ions. iv) high miniaturization and v) high sample throughput. The basic principle of operation of a DNA-chip consists of three main steps: 1) Functionalization.

Fig. discriminators and logic circuitry are all built onto the chip.3 Schematic diagram of an integrated DNA biochip system69 highly integrated biochips are made possible partly through the capability of fabricating multiple optical sensing elements and microelectronics on a single system.intensities of hybridization. In one biochip system. Most optical biochip technologies are very large when the excitation source and detector are considered. antibodies. such as DNA. making them impractical for anything but laboratory usage. a MFB which allows simultaneous detection of several disease end-points using different bioreceptors. Recently. An important element in the development of the multifunctional biochip (MFB) involves the design and development of an IC electro-optic system for the microchip detection elements using the complementary metal oxide silicon (CMOS) technology. while the latter shows the efficiency in rejecting analytes with any minimal mismatch68. amplifiers. each of the sensing elements is composed of 220 individual phototransistor cells connected in parallel to improve the sensitivity of the instrument. 70. The MFB device was a self-contained system based on an integrated circuit including photodiode sensor arrays. In this biochip the sensors. 3). amplifiers.2 Integrated DNA biochip The development of a truly integrated biochip having a phototransistor integrated circuit (IC) microchip has been reported by Vo-Dinh and coworkers69. Applications of the biochip are illustrated by measurements of the HIV1 sequence-specific probes using the DNA biochip device for the detection of a gene segment of the AIDS virus70. on a single biochip system was developed71. The ability to integrate light emitting diodes (LEDs) as the excitation sources into the system is also discussed. discriminators and logic circuitry. This work involves the integration of a 4 X 4 and 10 X 10 optical biosensor array onto an integrated circuit (Fig. electronics. 6. The multi-functional capability of the MFB biochip device is illustrated by measurements of different types of bioreceptors using . enzymes. With this technology. and cellular probes.

liquid chromatography-mass spectrometry (LC-MS) 75.infrared absorption spectrometry (LCIR)77. In some cases. expensive. 4 Schematic diagram of micro fluidics based biochip system71 spectrometry (MS.80. individual chromatographic-based separation systems generally have limited applicability in terms of ranges of molecular size and /or functionality. 4) 7. Furthermore. particularly with infrared. hyphenated techniques which couple spectroscopic techniques with separation systems are routinely used for the analysis of complex samples (e. etc.g.3 Micro fluidics-based biochip system The analysis of complex liquid samples for specific components necessitates some means of selective detection.g. and require skilled operators. Schematic diagram of micro fluidics based biochip system71 (Fig. liquid Chromatography . laborintensive. gas chromatography-infrared absorption spectrometry (GC-IR)73. Future issues in the development of Nanobiosensor and Biochip . ion trap MS79. and antibody probes targeted to the cancer related tumor suppressor gene p53. ion traps79. discrimination is afforded through spectral characteristics of the components in a complex sample. gas chromatography-mass spectrometry (GC-MS) 72-74.81-83. with the exception of some innovative MS device-based systems e. analysis of a complex sample may require multiple pretreatment steps as well as multiple chromatographic systems. 6. Raman and mass spectroscopes. Otherwise. 80.74. Even with hyphenated techniques.However. such instruments tend to be bulky. mass Spectrometry . capillary electrophoresis (CE).78.DNA probes specific to gene fragments of the Mycobacterium Tuberculosis (TB) system.77.MS)76.mass Fig.

Development of biosensor arrays that can successfully detect. blood analysis (see articles by Drs. Biochip technologies could offer a unique combination of performance . quantify and quickly identify individual components of mixed gases and liquid in an industrial environment. food and biomaterial quality testing. The size of device to move (levitate) tiny fluid droplet has been reduced by using micron scale diamagnets to create a magnetic micromanipulation chip. polymers. software. Biosensors offer several advantages over other analytical methods including rapid and even real-time measurements. plumbing. producing lab-on-a-chip devices (as described by Drs. It would be desirable to develop multiple integrated biosensor systems that utilize doped oxides. Biosensors are now being developed for detection of microbial pathogens and their toxins (see articles by Drs. selectivity. Even though micro array/biochip methods employing the detection of specific biomolecular interactions are now an indispensable tool for molecular diagnostics. Use of nanotubes. there are some limitations. In addition. The levitated particles can be manipulated and positioned with accuracy within a range up to 300 nm. Battrell and Gau and colleagues). The droplets used are 1 billion times smaller in volume than has been demonstrated by conventional methods.New biosensors and biosensor arrays are being developed using new materials. Buckminster fullerenes (buckyballs). silica and its derivatives can produce nanosized devices. Gau. and other physiological monitoring (see articles by Drs. and Sadik and their colleagues). biosensor technology is being applied to allergen detection. in the field or at the bedside. enzymes or other components to give the system the required specificity. Development of biosensors utilizing new technologies that offers improved sensitivity for detection with high specificity at the molecular level. As the technology advances. cancer detection (see articles by Drs. Use of this technology on a lab-on-a-chip would refine the examination of fluid droplets containing trace chemicals and viruses. There is also a need for reliable fluid handling systems for µdirty¶ fluids and for relatively small quantities of fluids (nanoliter to attoliter quantities). and specificity even when a complex or turbid sample matrix is used. A system with all the sensor components. and monitoring. Soper and Battrell and their colleagues). monitoring of glucose and other metabolites. high sensitivity. disposable. Gau. These should be low cost. 8. which operates with femtodroplets levitated in air84. Nanotechnologies can provide label-free detection and are being applied to overcome some of the limitations of biochip technology. nanomaterials and microfabricated materials including new methods of patterning. Thundat. Soper. Ligler. reagents and sample processing are an example of an integrated sensor. DNA micro arrays and enzyme-linked immunosorbent assay (ELISA) rely on the labeling of samples with a fluorescent tag²a procedure that is time consuming and expensive. Biosensor components will use nanofabrication technologies. and basic research on molecular interactions. these self-contained portable instruments will allow measurements outside the laboratory. and Battrell and their colleagues). Summary As biosensor technology advances. Yingfu Li and Homola and colleagues). reliable and easy to use as part of an integrated sensor system. Sensing in picoliter to attoliter volumes might create new problems in development of micro reactors for sensing and novel phenomenon in very small channels12. the range of applications broadens. Some of the challenges will be: Development of real-time non-invasive technologies that can be applied to detection and quantitation of biological fluids without the need for multiple calibrations using clinical samples.

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