Biosensor

Submitted by: Jyoti

10093004 MSc nanotech-4 sem

What is a Biosensor?

Biosensor Any device that uses specific biochemical reactions to detect chemical compounds in biological samples.

Current Definition
A sensor that integrates a biological element with a physiochemical transducer to produce an electronic signal proportional to a single analyte which is then conveyed to a detector.

Components of a Biosensor

Detector

Buffer flow

Valve

Membrane Biocomponent
Affinity Antibody Cell receptor Nucleic acid

Waste
Catalytic Enzyme Organelle Organism

Transducer
Electrochemical Optical Acoustics/Mechanical Calorimetric

Electrical Signal

Amplifier

Microelectronics

Data Processing

Father of the Biosensor

Professor Leland C Clark Jnr 19182005

History of Biosensors
1916 First report on immobilization of proteins : adsorption of invertase on activated charcoal 1922 1956 First glass pH electrode Clark published his definitive paper on the oxygen electrode.

1962
1969

First description of a biosensor: an amperometric enzyme electrodre for glucose (Clark)

Guilbault and Montalvo First potentiometric biosensor :urease immobilized on an ammonia electrode to detect urea Bergveld ion selective Field Effect Transistor (ISFET)

1970

1975 Lubbers and Opitz described a fibre-optic sensor with immobilised indicator to measure carbon dioxide or oxygen.

History of Biosensors
1975 First commercial biosensor ( Yellow springs Instruments glucose biosensor) 1975 1976 First microbe based biosensor, First immunosensor First bedside artificial pancreas (Miles)

1980
1982 1983 1984 detection

First fibre optic pH sensor for in vivo blood gases (Peterson)

First fibre optic-based biosensor for glucose First surface plasmon resonance (SPR) immunosensor First mediated amperometric biosensor: ferrocene used with glucose oxidase for glucose

History of Biosensors
1987 Blood-glucose biosensor launched by MediSense ExacTech

1990
1992 1996 1998 1998 FastTake

SPR based biosensor by Pharmacia BIACore

Hand held blood biosensor by i-STAT Launching of Glucocard Blood glucose biosensor launch by LifeScan

Roche Diagnostics by Merger of Roche and Boehringer mannheim

Current Quantom dots, nanoparicles, nanowire, nanotube, etc

Advances in several areas have led to development of Biosensors: Techniques for stabilizing enzymes/cells/Ab on to the surface of solid support while retaining their biological activity (Immobilization Technology) Development of tailor made membranes ( Membrane Technology) Integration of biomolecules and microelectronics/information technology (Semi conductor Industry)

Advantages of successful biosensor

Little sample consumption,high sampling rates, and relatively flexible and simple experimental set-up. Fast response time. Compactness due to miniaturisation. Parallel determination of more than one components. Reusability and disposability. Leads to predictable shelf life. Helps to achieve better quality and safety control,along with saving of time,energy,money and man-power.

Disadvantages of biosensor :
Limited availability. Limited testing life due to biological character of their design. Complexity and variability of the food samples have caused difficulty in their scale-up. Problems in mass production of biosensor components-immobilized biologically sensitive material and transducing system.

Basic Characteristics of a Biosensor

1. LINEARITY Linearity of the sensor should be high forthe detection of high substrate concentration. Value of the electrode response per substrate concentration. Chemicals Interference must be minimised for obtaining the correct result. Time necessary for having 95% of the response.

2. SENSITIVITY 3. SELECTIVITY

4.RESPONSE TIME

Biosensor
1. The Analyte (What do you want to detect)
Molecule - Protein, toxin, peptide, vitamin, sugar, metal ion

2. Sample handling (How to deliver the analyte to the sensitive region?)

(Micro) fluidics - Concentration increase/decrease), Filtration/selection

Biosensor
3. Detection/Recognition
(How do you specifically recognize the analyte?)

4. Signal
(How do you know there was a detection)

PRINCIPLE OF BIOSENSOR
A biosensor is an analytical measuring tool comprised of a biological element of known molecular properties tightly coupled to a physical transducer responsible for converting the biological signal into quantifiable information. Therefore, the biosensor selectivity is induced by the immobilization, in the sensitive area of the detector, of the biological component (enzyme, DNA receptor, antibody, antigen, microorganism, cell, etc.) specific to the target analyte. The molecular recognition then corresponds to the association of the biological element and its target molecule (analyte) through an association such as: enzyme-substrate, antibody-antigen, receptor-hormone, complementary DNA sequencing, etc. These associations maximize the capacity of the biomolecules to recognize a unique substance among various substances. The combinations of recognition-transducer systems are numerous and this explains the many definitions and nomenclatures of these types of sensors. The main methods of transduction that are the most current and well developed, from both a fundamental and experimental point of view, are: electrochemical, optical, acoustic and thermal.

COMPONENTS OF A BIOSENSOR

Physical components

Biological components

Physical components (i) TRANSDUCER A transducer converts the biochemical signal to an electronic signal. The signal produced may be in the form of electrochemical (change in potential or current), optical (color change), calorimetric (heat measurement), piezoelectric (mass change) response etc.

(ii) AMPLIFIER The electronic signal produced by transducer is very small & is amplified by an amplifier. (iii) MICROPROCESSOR The amplified signal is fed in to the microprocessor. The signal is then processed & interpreted & is displayed in suitable units.

Biological components
Enzymes These may be used in purified form or may be present in microorganisms or in slice of intact tissue. They are biological catalyst for particular reactions and can bind themselves to specific substrate. This catalytic action is made use of in the biosensor. Antibodies They will bind specifically with the corresponding antigen. They have no catalytic activity. Despite this, they are capable of developing ultra high sensitivity in biosensors. Nucleic acids They operate selectively because of their base pair characteristics. Receptors Inside the lipid bilayer plasma membrane surrounding a cell are protein than transfer the full breadth of the membrane and which have molecular recognition properties. They are difficult to isolate but will bind solutes with high degree of affinity and specificity matching antibodies.

The biological component interacts specifically to the analyte, which produces a physical change close to the transducer surface. This physical change may be: Heat released or absorbed by the reaction (measured by calorimetric biosensor). Production of an electrical potential due to changed distribution of electrons (potentiometric biosensors). Movement of electrons due to redox reaction (amperometric biosensor). Light produced or absorbed during the reaction (optical biosensor).

OTHER COMPONENTS OF A BIOSENSOR SYSTEM

ANALYTE/SUBSTRATEf: It is compound whose conc. is to be determined by the biosensor, either by being consumed or produced in a biochemical process. Among the analytes detectable with biosensors are carbohydrates, organic acids, alcohols, phenols, amines, heterocyclic compounds, proteins, enzymes cofactors, inorganic ions, hormones, vitamins, peptides, drugs, xenobiotics, microorganisms, gases, and pesticides. Analyte concentrations measurable with various types of biosensors range rom 10-7 molL-1 (biocatalytic sensor) to as little as 10-15 molL-1 (affinity sensors). MEMBRANES: A membrane that is selectively permeable normally protects the receptor. The art of membrane construction, though relatively new, is rapidly evolving to meet the needs of commercial biosensor construction. It is highly specialized, involving techniques from simple solution casting to spin- and spray- coating, and interfacial polymerization together with electropolymerisation at a surface.

CLASSIFICATION OF BIOSENSORS
Based on the level of integration Based on transducer system Based on sensing element

inBased on the level of integration

FIRST GENERATION BIOSENSORS In case of these biosensors, the biocatalyst is either bound to or entrapped in a membrane, which in turn is fixed on the surface of the transducer. SECOND GENERATION BIOSENSORS These biosensors involve the adsorption of covalent fixation of the biologically active component to the transducer surface. THIRD GENERATION BIOSENSORS In these biosensors the biocatalyst is directly binded to an electronic device that transduces and amplifies the signals.

BASED ON TRANSDUCER
ucer system Electrochemical biosensors Optical biosensors Calorimetric biosensors Piezoelectric biosensors Thermal biosensors

Electrochemical biosensors
The biological signal can be used to generate a current or charge or may change conductivity between two electrodes and therefore the corresponding transduction device has been described as Amperometric, Potentiometric Conductometric

Optical biosensors
Optical biosensors are based on the measurement of the light absorbed or emitted as consequence of a biochemical reaction. In such type of biosensors, light waves are guided by means of optical fibers to suitable detectors. These types of biosensors have been used for the detection of pH, O2 and CO.

Calorimetric biosensors
The basic principle of such biosensor is that all biochemical reactions is involve a change in enthalpy such a change in enthalpy is detected by calorimetric biosensors for detection, Thermal signal generated by the redox reaction was measured as opposed to measuring the electrochemical signal.

Piezoelectric biosensors
These biosensors operate on the principle of generation of electric dipoles on subjecting anisotropic natural crystal to mechanical stress. Such type of biosensors utilized for the measurement of NH3, H2, CH4, CO & other organophosphorous compounds. This is also called as acoustic wave biosensor.

Thermal biosensors
Thermometric devices predominantly measure the changes in temperature of the circulating fluid following the reaction of a suitable substrate with the immobilized enzyme molecules.

 Catalytic Biosensors Enzyme and microbial cell based biosensors come in this category. The immobilized sensing biomolecule act as a catalyst and catalyze the biochemical reaction. Various examples of catalytic biosensors are Glucose Biosensor (glucose oxidase based), Cholesterol biosensor (cholesterol oxidase and cholesterol esterase based), Urea biosensor (urease based), Lactate biosensor (lactate oxidase and lactate dehydrogenase) etc. Affinity Biosensors This type of biosensors exploits the affinity between the analyte and biocomponent immobilized onto the sensor surface. Biomolecules such as antibodies, nucleic acids, lectins and hormone receptors, are used to bind complementary molecules irreversibly and non-covalently. The basic principle behind the affinity biosensors is that the analyte must be the complementary to the immobilized biocomponent. Immunosensors and DNA biosensors are usually the most studied in this category.

Based on sensing element

MATRIX FOR BIOSENSOR FABRICATION

A number of matrices have been used for the immobilization of biomolecules to improve their activity and stability. Example: - Conducting polymers. - Sol-gel matrices. - Nanomaterials. - Composites of nanomaterials and conducting polymer.

Methods of immobilization
Adsorption Microencapsulation Entrapment Covalent attachment Cross linking

Advantages of biosensors
High selectivity and specificity High sensitivity Fast respose time Simple to use Comparatively low operation cost Miniaturization Disposibility

Disadvantages of biosensors
Limited availability Instability Limited shelf life Sensitivity to interference and contamination

Applications
Clinical diagnosis and biomedicine Food and drink production and analysis Pharmaceutical and drug analysis Industrial effluent control Pollution control and monitoring Mining, industrial and toxic gases Military applications Farm, garden and veterinary analysis Process control: fermentation control and analysis Microbiology: bacterial and viral analysis

Applications of Biosensors

Continuous monitoring Sensitive Detection of nutrients/other parameters Can be incorporated into Products to monitor
Temperature abuse Shelf life Contamination, and To provide visual indicator to consumers of the state of the product at the time of purchase

Commercially available Biosensors for Food Analysis Analyte Lactose Sucrose Lactate Ethanol Amino acids Biocomponent -Galactosidase Invertase Lactate DH ADH Amino acid DH Raw milk Instant cocoa manufacture Dairy products, Yoghurt, whey Alcoholic beverages Many foods Application

Glutamate
Ascorbic acid Pencillin PHB ester Sulfite Aspartame Glucose

L- Glu oxidase
Ascorbate oxidase Ab- enz conjugate P- OH Benzoate hydroxylase Sulfite oxidase L-aspartase/ Al. oxidase Glu oxidase

Soy sauce manufacture

Fruit juices Milk Fruit juices & drinks Dry fruits, wine, vinegar. Many foods (level of sweetener) Fruit juices, molasses, Brewing etc

ELECTRONIC TONGUE

Its applications include: Bitterness Measurement of New Chemical Entities Bitterness Masking Efficiency Matching Placebo Development (same taste of drug) Shelf-life & Stability of Formulations Comparison with Competitor Products Scale-up of Formulation Process Screen for best Additive & Taste agents

ELECTRONIC NOSE The US Department of Agriculture (USDA) is testing the zNose technology to see whether the sensor devices can pick up bug infestation in plants. The zNose could have a wider use for food and beverage companies, such as for aroma chemistry testing or for the analysis of wine and beer. The zNose is unique in its ability to separate and quantify the chemistry of any fragrance, odor or chemical vapor with part pertrillion sensitivity within 10 seconds.

 A researcher examines a capillary column used as an electrochemical sensor to detect harmful bacteria in food products. Such sensors promise to speed up detection of harmful bacteria in food processing industries.

Electroimmunoassay technology is composed of a circuit with a capture antibody attached to the solid surface in the area of the electrode gap

HAND HELD MODEL OF A BIOSENSOR

Using a DNA-based cell adhesion system, researchers have created cell chips (self-assembled arrays of cells on a thumbnail-sized chip), analagous to DNA chips, that could be used as biosensors for detecting the presence of pathogens.

A new technique in which single strands of synthetic DNA are used to firmly fasten biological cells to non-biological surfaces has been developed by researchers with the Lawrence Berkeley National Laboratory (Berkeley Lab) and the University of California at Berkeley.

BIOSENSORS FOR PACKAGES ARE BEING DEVELOPED WHICH CAN NOT ONLY INDICATE THE FRESHNESS OF FOODS INSIDE THE PACKAGE BUT ALSO DETECT THE PRESENCE OF PATHOGENIC MICROORGANISMS AND THEIR TOXINS

Work is being done on RFID biosensors, which could be implanted into body to monitor the body functions of patients

A pateints left hand with the planned location of the RFID chip

BIOSENSOR USED FOR ENVIRONMENTAL MONITORING

New biosensor device developed by University of Rhode Island to detects Salmonella, E. coli, and other bacteria. The biosensor uses fiber optic technology to quickly and accurately detect and quantify bacteria levels in meats, poultry and other foods. The latest version also uses microscopic magnetic beads called microspheres. The surface of the beads are covered with antibodies that collect the pathogen and are then labeled with a fluorescent dye. Then the beads are magnetically focused in front of optical fibers and a laser signal reports the pathogen concentration.

Wearable Biosensors
Ring Sensor

Smart Shirt

Biosensors on the Nanoscale

Molecular sheaths around the nanotube are developed that respond to a particular chemical and modulate the nanotube's optical properties. A layer of olfactory proteins on a nanoelectrode react with low-concentration odorants (SPOT-NOSED Project). Doctors can use to diagnose diseases at earlier stages. Nanosphere lithography (NSL) derived triangular Ag nanoparticles are used to detect streptavidin down to one picomolar concentrations. The School of Biomedical Engineering has developed an antibody based piezoelectric nanobiosensor to be used for anthrax,HIV hepatitis detection.

Thank you!