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    BIOSENSORS

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    glorynjoku20

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    © All Rights Reserved
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    BIOSENSORS

    Uploaded by

    glorynjoku20
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    BIOSENSORS

    Uploaded by

    glorynjoku20

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    © All Rights Reserved
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    BIOSENSORS 1

    BIOSENSORS

    1 BIOSENSORS
    Introduction: The first biosensor was described by Clark and Lyon in 1962. They described a
    sensor consisting of glucose oxidase immobilised on an amperometric oxygen electrode
    intended for measuring the amount of glucose in solution.

    1.1 IUPAC Definition of a Biosensor:


    A self-contained integrated device which is capable of providing scientific quantitative or
    semi-quantitative analytical information using a biorecognition element (biochemical
    receptor) which is in direct spatial contact with a transducer element.

    Figure: Biosensor schematic

    1.2 Components of a Biosensor


     Bioreceptor: immobilised, sensitive biological element e.g. enzyme, DNA, probe,
    antibodies) able to recognise analytes.
     Transducer: converts (bio)chemical signal resulting from analyte:receptor interaction into
    an electronic signal whose intensity is either directly or inversely proportional to analyte
    concentration. Electrochemical transducers are most common because they are cheap,
    have a simple design and usually very small. Other types of transducers include
    gravimetric, calorimetric, optical detection.

    1.3 Types of Biosensors


    Categories of biosensors are based on their principles of signal transduction and
    biorecognition element. The following types are common

     Electrochemical
    o Potentiometric
    o Amperometric
    o Conductimetric
     Optical
     Resonant
     Piezoelectric
     Thermal
     Ion sensitive

    1.4 Advantages of using biosensors for diagnostic (and other) purposes


    1. They have the ability to measure non-polar molecules which would have otherwise been
    difficult to measure

    BIOSENSOR MLS 402


    ELECTROCHEMICAL BIOSENSORS 2
    BIOSENSORS

    2. They are specific due to the immobilised biorecognition element


    3. Rapid and continuous control of measurement is possible
    4. Short response times (typically less than a minute)
    5. Very robust and practical.

    1.5 Disadvantages
    1. Heat sterilisation is not possible (biologicals are easily denatured)
    2. Stability of biologicals used is not always guaranteed, they are prone to environmental
    effects (heat, pH, humidity etc.)
    3. They are prone to interference and most times destruction of cells (if used in biosensing)
    by other molecules in the measured analyte.

    1.6 Considerations in Developing a Biosensor


    1. Choose a suitable biomolecule (sensing element)
    2. Choose suitable immobilisation method
    3. Choose appropriate transducer
    4. Be conversant with all the basic biological, physical, kinetic and chemical principles
    covering all elements you intend to use.
    5. Work with other professionals, it is an interdisciplinary venture.

    2 ELECTROCHEMICAL BIOSENSORS
    These have been developed to be very specific, rapid, sensitive, highly selective and cheap (in
    principle) for analysing bio species. They find applications in clinics, process control
    (industries and environment), in vivo studies (implantable sensors), food industry, security
    (bio warfare).

    The investigated biochemical reaction would usually generate measurable current


    (amperometric), potential/EMF/charge accumulation (potentiometric) or conductivity
    (conductimetric) changes between electrodes. Another variant makes use of measured current
    with controlled variations of applied potential and this is called voltametric.

    Biologicals are not normally electroactive so their combination with the biorecognition
    element is coupled to reactions that produce an electroactive element which can be detected.

    Electrochemical cells consists of three kinds of electrodes

     Working electrode: where the reaction occurs. E.g. include glassy carbon electrodes,
    screen printed electrodes, gold, silver, tin electrodes. Screen printed electrodes are made
    by depositing thin films of electrodes on substrates. They are usually cheap and
    disposable. The working electrodes may be anodic or cathodic depending on the set up.
     Reference electrode: may be hydrogen, calomel or glass. Other electrodes “refer” to this
    one for their function. Types include Ag/AgCl in NaCl solution, calomel. Measurements
    are taken at the working electrode with reference to this electrode.
     Auxiliary electrode: also called counter electrode, it complements the working electrode
    (e.g. cathode when working electrode is anode and vice versa).

    BIOSENSOR MLS 402


    FIBRE OPTIC BIOSENSORS 3
    BIOSENSORS

    Figure: Schematic representation of a measuring cell.

    2.1 Factors Affecting Electrochemical Biosensors


    Selectivity, response time, sensitivity range, accuracy, recovery time, solution, life span.

    2.1.1 Immobilisation on Working Electrodes


     Adsorption (physical, chemical). Initial microencapsulation using cellulose, collagen
    improves adsorption.
     Entrapment
     Covalent attachment (as long as the active groups remain available)

    3 FIBRE OPTIC BIOSENSORS


    Small flexible wires made of glass or plastic able to transmit light over long distances without
    appreciable loss of signal. FOBs are good for harsh environments because they are strong,
    flexible and cheap. A major attraction to optical fibres is that they can permit the transmission
    of multiple signals synchronously giving it the ability to sense multiple analytes.

    Optical fibres usually consist of a core and a coating (cladding/jacket) both having different
    refractive indices (n1 and n2). These refractive indices interfere with each other forming a
    mirror because they are different. This leads to a series of internal reflections from end to end
    (total internal reflection).

    Figure: Fibre optic cable illustrating total internal reflection

    Fibre optic biosensors generally consists of

     Light source (tungsten lamp, LEDs, xenon lamps, laser)


     An optical fibre
     Sensing material
     Detector (photomultiplier, CCDs)

    Reaction between sensing element and analyte leads to a change in both physicochemical and
    optical properties. The transduction mechanism produces optical signal related to analyte

    BIOSENSOR MLS 402


    PIEZOELECTRIC BIOSENSORS 4
    BIOSENSORS

    concentrate. Measuring optical signal involves measuring difference between incidence and
    output light at the location where the sensing element has been immobilised. Output light is
    sent to the detector by the fibre (either reflected, emitted or absorbed light is measured).

    3.1 Advantages
    1. No reference electrode is needed
    2. The sensors are very mobile since no extra reagent comes in contact with the FOB
    3. No electrical hazard or interference
    4. There is less dependence on temperature
    5. Easy to miniaturise hence has value for micro measurement
    6. Multiple analytes can be measured once the wave forms are guided
    7. Can theoretically be used for most chemical analytes

    3.2 Disadvantages
    1. Short life span of reagents
    2. Analyte diffusion leads to longer response time
    3. Needs special reagents
    4. Not easily commercialised.

    3.3 TYPES OF FOBs


    1. Absorbance FOB – detects transmitted or scattered light obeying Beer-Lambert’s law
    2. Fluorescent FOB - more commonly used because of high sensitivity even at small
    concentrations. The transmitted light excites the fluorophore leading to fluorescence
    which is measured by a detector
    3. Luminescence FOB – may be chemiluminescence or bioluminescence. Chemical or
    biological reaction leads to emission of light. Bioluminescence occurs naturally. Luciferase
    enzyme system is widely used.
    4. Reflectance FOB – works with evanescent waves. Reflection of light changes due to the
    biorection (and change in structure of the structure of the bio-recognition elements).

    4 PIEZOELECTRIC BIOSENSORS
    These biosensors make use of piezoelectric crystals which vibrate under the influence of an
    electric field. E.g. of such is Quartz crystals. The initial frequency of vibration is characteristic
    of such crystals and usually depends thickness and cut of the crystal. This resonant frequency
    changes as the crystals adsorb or release adsorbed molecules from their surface. The change
    in resonant frequency is usually directly proportional to the change in adsorbed mass within
    the limit of a small percentage of change.

    These changes in resonant frequency are then detected by special electronic circuits. An
    example is the formaldehyde biosensor which uses crystal-immobilised formaldehyde
    dehydrogenase to detect formaldehyde gases.

    4.1 Advantages
     They are inexpensive, robust and small
     They give a rapid response.

    4.2 Disadvantages
     There is pronounced interference from atmospheric humidity.

    BIOSENSOR MLS 402


    RESONANT BIOSENSORS 5
    BIOSENSORS

     They cannot be used to detect samples in solution.

    5 RESONANT BIOSENSORS
    This makes use of an acoustic wave transducer coupled to a membrane bound bio recognition
    element. On interaction of the analyte and bioelement, there is a change in mass of the
    membrane changes. The change in mass induces a measurable change is the resonant
    frequency of the wave transducer.

    6 THERMAL BIOSENSORS
    These sensors leverage on thermal changes (absorption or production of heat) inherent in
    biological reactions. These changes in turn affect the temperature of the medium in which the
    reaction occurs.

    These sensors are designed by immobilising enzyme molecules with temperature sensors
    such that when the analyte reacts with the enzyme, the heat reaction of the enzyme is
    measured and calibrated against the concentration of the analyte.

    In ideal situations, total change in temperature is directly proportional to the molar enthalpy
    (i.e. change in temperature when 1 mole of a substance is undergoing change) and total
    number of molecules taking part in the reaction. Measurements are taken using a thermistor
    (a kind of resistor whose resistance decreases with increasing temperature).

    They are mostly used in pesticide and bacterial detection

    6.1 Advantages
     They are very sensitive and do not require constant calibrations.
     They are insensitive to optical and electrochemical properties of the samples hence less
    interference.

    7 ION SENSITIVE BIOSENSORS


    These are semiconductor field effect transistors (FETs) having an ion-sensitive surface. The
    surface electrical potential changes when the ions and the semiconductor interact. This change
    in the potential can be subsequently measured. The Ion Sensitive Field Effect Transistor
    (ISFET) can be constructed by covering the sensor electrode with a polymer layer that is
    selectively permeable to analyte ions. The ions diffuse through the polymer layer and in turn
    cause a change in the FET surface potential. This type of biosensor is also called an ENFET
    (Enzyme Field Effect Transistor) and is primarily used for pH detection.

    BIOSENSOR MLS 402

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