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420 ANALYTICAL ASPECTS of FET-BASED BIOSENSORS Jing-Juan Xu, Xi-Liang Lu

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420 ANALYTICAL ASPECTS of FET-BASED BIOSENSORS Jing-Juan Xu, Xi-Liang Lu [Frontiers in Bioscience 10, 420--430, January 1, 2005] ANALYTICAL ASPECTS OF FET-BASED BIOSENSORS Jing-Juan Xu, Xi-Liang Luo and Hong-Yuan Chen Key Lab of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093, P.R. China TABLE OF CONTENTS 1. Abstract 2. Introduction 3. Basic structure and principle of bioFET 4. Immobilization of biological components 5. Classification and application of bioFETs 5.1. Catalytical BioFETs 5.2. Affinity BioFETs 5.3. Cell-based BioFETs 6. Development of bioFETs 6.1. Improvement on configurations of ISFET-based biosensor 6.2. Application of nano materials to bioFETs 7. Summary and perspective 8. Acknowledgement 9. References 1. ABSTRACT Field-effect transistor (FET)-based biosensors response and high sensitivity; (e) low-cost mass production. (BioFETs) have undergone great progress especially in the Therefore, these advantages made ISFET an ideal device last decade, since they were first realized in 1980. Recently, for the construction of biosensors, especially portable micro BioFETs have become one of the most important branches biosensors. Since the first report of BioFET by Caras and of biosensors. This paper briefly reviewed the operating Janata in 1980 (1), a great number of publications have principles of BioFETs and summarized the improvement been devoted to this field, and a few useful reviews (2-4) and application of BioFETs, finally, the future prospects of have partly summarized these works (mainly published in BioFETs were discussed with 126 references. 1995-2001). With the development of science and technology, some important works and new methods 2. INTRODUCTION involving BioFETs have been reported in succession. This paper gives a brief introduction of the principle of In recent years, research in the field of biosensors BioFETs, and mainly focuses on the application and recent has received great attention because of their extensive improvement of BioFETs. application in agriculture, environment, clinical analysis and food industry. Biosensors are normally composed of 3. BASIC STRUCTURE AND PRINCIPLE OF two main parts: the biological recognition part and the BIOFET signal transducing one. As functional hybrid systems, they benefit from the coupling of the unique recognition and BioFETs are mainly composed of two parts: the signal-amplification abilities of biological systems. biological recognition element and the ISFET. The Therefore, the integration of biologically active materials biological recognition element is the sensing part that together with different types of transducers offers the immobilized with biological materials, such as enzyme, possibility of the generation of highly sensitive, specific, cell, DNA etc., while the ISFET acts as the signal selective and reliable biosensors. Among various types of transducing part. Among the various kinds of ISFETs used transducers used for biosensors, ion-sensitive field-effect for BioFETs, the most common and useful one is the H+- transistor (ISFET) is one of the most attractive selections. sensitive field-effect transistor. The basic structure of an The ISFET was first developed in the early 1970s and has ISFET is the ordinary metal-oxide-semiconductor field- been introduced as an alternative to the fragile glass effect transistor (MOSFET) with the metal gate replaced by electrode for the measurement of pH and other ions an ion-sensitive membrane, an electrolyte solution and a concentrations. Despite the inherent problems, such as reference electrode. It consists of a silicon substrate with temperature sensitivity, light sensitivity and drift, the two doped regions, source and drain, separated by a ISFET has received increasing attention in the construction channel that under the gate insulator. In the case of a of ISFET-based biosensors (BioFETs). The silicon-based BioFET, biological recognition material is immobilized on ISFET has a lot of significant characteristics and benefits: the gate insulator surface to form the sensing membrane. (a) high input impedance and low output impedance; (b) The gate insulator is typically a SiO2 layer or a SiO2 layer small size and weight; (c) anti-breaking ability: the all- covered with another layer of Si3N4, Al2O3 or Ta2O5. The solid-state ISFET is very robust and durable; (d) fast upper layer of the gate insulator structures serve as 420 Analytical Aspects of FET-based biosensors Figure 1. Equivalent circuit of an ISFET device modified by a membrane on the gate interface.(from Ref. 8 and 10). + sensitive materials for H -sensitive ISFETs, and the where pHpzc is the value of the pH for which the oxide sensitivity of these materials is in the order of SiO2 < Si3N4 surface is electrically neutral; β is a parameter which < Al2O3 < Ta2O5 (5). reflects the chemical sensitivity of the gate insulator and is dependent on the density of surface hydroxyl groups and For operating an ISFET, the gate voltage, VGS, is the surface reactivity; k is the Boltzmann constant and T is applied by a reference electrode through the electrolyte the absolute temperature. Thus, by measuring changes in solution. When a sufficiently positive bias potential is the drain current, the pH value of the test solution can be applied to the gate, an inversion layer is induced in the determined. In most cases, ISFETs are often operated in the channel between source and drain. The magnitude of the constant drain current mode, that is, setting the drain drain current, ID, will be determined by the effective current at a fixed value by a feedback circuit, and electrical resistance of the surface inversion layer and the measuring the voltage shift that results from the reaction voltage that is applied between source and drain, VDS (3). near/at the gate (sensing membrane). Since the channel resistance in ISFET depends on the electric field perpendicular to the direction of the current, Besides these methods involving the charges from solution accumulate on top of the gate measurement of current and voltage, another technique, insulating membrane influence the interfacial potential and impedance spectroscopy was also introduced to the analysis then the drain current. The dependence of the interfacial of biorecognition processes at the gate surface of the ISFET potential on the charge concentration can be explained by by Willner and coworkers (8, 9) recently. The impedance the so-called site-binding theory (6). Generally, the of the membrane associated with the ISFET can be operational mechanism of the ISFET can be described by represented by the equivalent circuit shown in Figure 1 (8, an expression of the drain current, ID, in the unsaturated 10). Where RSi, Rmem, Rct, and Rsol correspond to the region (3, 5, 7): resistances of the Si layer, the chemical sensing interface, [1] the space charge, and solution, respectively; Csc, Cox, Cmem, W Φ Q + Q Q V 2 I = µC V − E − Ψ + χ sol − Si − OX SS − B + 2φ V − DS D OX GS ref 0 f DS and Cdl correspond to the space charge capacitance and the L q COX COX 2 capacitances of the oxide insulating layer, the membrane, where C is the capacitance of the gate oxide; W and L are ox and the double layer, respectively; W represents the respectively the width and the length of the channel; µ is Warburg impedance. If a biorecognition process such as the the average electron mobility in the channel; E is the ref formation of antigen-antibody or hybridization of DNA potential of the reference electrode; V and V are GS DS occurs, the correlated changes in the impedance will be respectively the gate source voltage and the drain source caused. By monitoring the changes in impedance, the voltage; Ψ is the electrostatic potential; χsol is the surface 0 amount of antigen/antibody or complementary DNA (11) dipole potential of the solvent; Ф is the silicon electron Si can be determined. work function; q is the elementary charge; QB is the depletion charge; Qox and Qss are the charges located in In brief, the ISFET is very sensitive to electrical the oxide, and the charges located in surface states and interaction at or near the gate insulator/electrolyte interface, interface states respectively; ф is the potential difference f and therefore each biochemical reaction/process leading to between the Fermi levels of doped and intrinsic silicon. For chemical or electrical changes at this interface can be a given ISFET, all parameters are constant except Ψ and 0 measured by the BioFET immobilized with different χsol. In the case of a H+-sensitive ISFET, the surface dipole biological recognition materials. potential, χsol, is supposed to be independent of pH and therefore, changes in the drain current are attributed to 4. IMMOBILIZATION OF BIOLOGICAL changes in the electrostatic potential, Ψ , only. According 0 COMPONENTS to the site-binding theory, the electrostatic potential of a H+-sensitive ISFET can be given as follows (3, 5): The crucial procedure for the construction of kT Ψ = 2.3( )[β /(β +1)]( pH − pH ) [2] biosensors, including BioFETs, is the immobilization of 0 q PZC biological component. A number of methods for the 421 Analytical Aspects of FET-based biosensors biological components immobilization have been and poly(4-vinylpyridine-co-styrene) (28, 29, 39, 40, 43) developed. have been applied on top of the enzyme layer to control the diffusion of substrate and product. With this approach, the 1. Physical or chemical adsorption (12-14), which is the dynamic range of glucose ENFET can be extended up to 20 simplest immobilization method. mM (28). An additional Pt electrode is deposited on the 2. Cross-linking (15-19). A bi- or multi-functional reagent gate region of glucose ENFET to electrolyze hydrogen is employed to form a network-like structure between peroxide and generate two hydrogen ions (15, 34), and the biomolecules, biomolecule and gel/polymer matrix. sensitivity of the glucose ENFET improved four times. 3.
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