Bioelectronic Devices:Biosensors

What are Biosensors?

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Biosensors are small analytical devices which identify the analyte and modify it into a product detectable by a transducer which transforms it into an electrical or electronic signal.

PrinciplesofBiosensors

The first link of a biosensor is the bioreceptor, which has a particularly selective site that identifies the analyte(substrate). The bioreceptor (biologically active substance) ensures molecular recognition and then transforms it or modifies it into a product detectable by a transducer. This modification of substrate by the receptor is a biochemical modification producing a physicochemical effect. Transducer then transforms this modification into an electrical or electronic signal.

Biosensor

EnzymesElectrodes

microorginismsTransistors
AntibodiesThermistorsAmp

Receptorsopticalfibers
CellsandTissues

SampleBioreceptorTransducerSignal

Schematicrepresentationofbiosensor

Transduction mechanism of biosensors: thetotaleffectofabiosensoristotransforma

biologicaleventintoanelectricalorelectronicsignal. Bio-chemical signal S Binding sites Biological membrane

H+ n+ n+

P-Si B

Bioreceptors

Bioreceptor is made of highly specialized macromolecules with the appropriate specificity and sensitivity. The following biocomponents are generally used as
BIORECEPTOR

Different Bioreceptors
Enzymes: These are proteins that lower the energy threshold at which a given reaction takes place.A large number of enzymes are commercially available, for example, glucose oxidase and urease. s Antibodies(Immunoreceptors): The antibodies are glycoproteins produced by the immune system
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processes at the molecular level is based on specialized protein structures, called receptors, able to recognize a number of physiological signals. The acetylcholine receptor is the best known receptor in the field of neurotransmission.

Receptors:The regulation of biological .

Cells and Tissues: If a biosensor makes use of complex

reaction schemes then it may be more convenient to use microorganisms or cells where the metabolic pathway is naturally present. Tissue is also another option, has the advantage of cohesion, has a structure that is robust enough to be attached directly to the transducer.

Biodetection Techniques
A natural event is characterized by certain properties, which can be exploited for its identification using biodetection methods.The senses of living organisms are the most apt tools for the specific recognition of these properties. Out of our five senses, sight, touch and hearing exploit preferentially the principles of physics, whereas smell and taste make more use of chemical reactions.Thus in living organisms, the information is transmitted through an assembly of physicochemical systems.In reality, biosensor is also an assembly of physicochemical systems. In a biosensor, the transducer plays

Electrochemical Detection (a) Amperometric techniques: This technique measures the intensity of current,generated by oxidationreductions induced by the biological component, toward a fixed electrode. Oxidation or reduction of a species is performed by a working electrode, and a second electrode acts as a reference.Example:-Glucose

Glucose oxidase Glucose +O2 Gluconic acid + H2O2 +0.65V H2O2 2H+ + O2 +2eFig(a): Reactions in a glucose sensor Here either the H2O2 or the amount of oxygen consumed during the oxidation of glucose can be detected. In the case of H2O2, during electrolysis ,the working electrode(platinum) acts as an anode because it is polarized to a positive potential +0.65 V

(b) Potentiometric technique:

This technique measures the difference in potential between two electrodes immersed in solution.One of the electrodes probes the solution, while the other serves as the reference. The reference electrode potential is independent of its environment while the potential of the probe electrode is the potential at the interface between the solid and liquid phases.Glass pH electrode, MOSFET sensors are based on this technique.

Thermometric Detection This method detects the variations in enzymatic reaction enthalpies.Thermistors or thermopiles are used to detect small variations in temperature.

Photometric Detection This method uses the principle of emission, absorption or scattering of light and makes large use of optical fibers

Classification of Biosensors

Biosensors can be classified according to the Bioreceptor used for detection and sensing o can be classified according to the type of transducers used.Table 1 shows the classification of biosensors with an idea of the approximate state of research as a function of the different possibilities of combination between various bioreceptors and transducers.

Bioreceptor/biosensor Enzyme/Enzymatic
XXX XXX XX X X X

Transducer

Amperome Electroch tric emical Potentiome cell tric ISFET Thermometric Optical Piezoelectric

Antibodies / Cell/Cell Tissue/Tissue immuno

XX XX X X X X XX XX X X

Table1:ClassificationofBiosensors X:Stateoffundamentalresearch XX:ResearchandDevelopmentofprototypes XXX:ResearchandcommercialDevices

Bioelectronic sensor

ISFET:
device

Transducer for Bio-electronic

The Ion-Sensitive Field Effect transistors (ISFETS) are sensors to measure the level of specified ions in aqueous solution. In the case of ISFET the metallic gate of MOSFET is replaced by a specific oxide-coated gate,that is sensitive to hydrogen ion concentration. When immersed in a liquid, the electrical circuit Vgs is closed with the reference electrode and the hydrogen ion concentration in solution can influence the drainsource current. Ions or other charged particles do not have to pass through the ion sensitive membrane. The control of drain current is solely based on electrostatic effects. The interaction between the ions and the thin film changes the properties of the insulator -electrolyte interface, which leads to a modification of the threshold voltage, and hence a change in the drain current.

MOSFET to ISFET

What is BIOFET?
The biosensor which uses ISFET as a transducer is called BIOFET ENFET: Glucose sensor, Urea sensor, Creatinine sensor etc. s IMMUNOFET: s DNAFET
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What is ENFET An ENFET is a biosensor which uses enzyme as bioreceptor. It is constructed by placing an enzymatic membrane over the ion-selective membrane of an ISFET. The product of the enzymatic reaction chosen for measurement is detected by Reference electrode the ISFETVG
Enzyme SiO2

VD nS
P-S i

nD ID

Principle of operation

The sensitive surface of the transducer is in contact with the enzymatic laye The external surface of the enzymatic layer is immersed in the solution containing the substrate.

Transducer (ISFET)

enzyme S P

S solution

Figure: Schematic representation of the diffusion of the substrate S and the product P in the enzymatic layer on a transducer.

Steps required during the operation:

transport of the substrate from the bulk of the solution towards the zymatic layer.

Diffusion of the substrate within this layer, accompanied by the zymatic transformation of the substrate into reaction product.

Migration of the product towards the transducer,

conversion of the concentration of the product at the interface of e enzymatic layer and transducer into an electrical signal by the transdu

Theoretical aspects

The presence of the enzyme ensures the transformation of the subst into the reaction product according to the following reaction: k+1 k+2 E+S ES E + P k-1 where E represents the enzyme, S the substrate and P the reaction product; k+1, k-1 and k+2 are the rate constants of the reactions.

One example

Glucose oxidase catalyses the transformation of glucose into gluconolactone which in turn hydrolyses into gluconic acid. Due to acid formation, the local pH changes which can be detected by the adjacent ISFET. Glucose oxidase Glucose + O2 + H2O H2O2 gluconic acid

Some basic enzyme reactions which are used for ENFET creation
H2N Urease C = O + 2H2O + H+
H 2 2NH

4+ + HCO3-

N CD N-methylhydantion + NH4+

Creatinine + H2O

CD is the creatinine deiminase

These reactions generate or consume protons, changing the pH inside the enzyme membrane, which is registered by the ISFET

Figure: Schematic representation of ENFET from R&D Institute of Microdevices (Kiev,Ukraine).