microbial detection using biosensor. what is biosensor ? analytical device which utilize biological...
TRANSCRIPT
![Page 1: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/1.jpg)
MICROBIAL DETECTION USING BIOSENSOR
![Page 2: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/2.jpg)
WHAT IS BIOSENSOR ?
Analytical device which utilize biological reaction of biochemical molecule for detecting
target analyte and converts a biological response into a quantifiable and processable
signal
Pregnancy test
Glucose monitoring device (for diabetes patients)
![Page 3: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/3.jpg)
“Biosensor” – Any device that uses specific biochemical reactionsthat uses specific biochemical reactions to to
detect chemical compoundsdetect chemical compounds in biological samples.
![Page 4: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/4.jpg)
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.
![Page 5: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/5.jpg)
Father of the Biosensor
Professor Leland C Clark Jnr 1918–2005
![Page 6: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/6.jpg)
1916 First report on immobilization of proteins : adsorption of invertase on activated charcoal
1922 First glass pH electrode
1956 Clark published his definitive paper on the oxygen electrode.
1962 First description of a biosensor: an amperometric enzyme electrodre for glucose (Clark)
1969 Guilbault and Montalvo – First potentiometric biosensor:urease immobilized on an ammonia electrode to detect urea
1970 Bergveld – ion selective Field Effect Transistor (ISFET)
1975 Lubbers and Opitz described a fibre-optic sensor with immobilised indicator to measure carbon dioxide or oxygen.
History of Biosensors
![Page 7: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/7.jpg)
1975 First commercial biosensor ( Yellow springs Instruments glucose biosensor)
1975 First microbe based biosensor, First immunosensor 1976 First bedside artificial pancreas (Miles) 1980 First fibre optic pH sensor for in vivo blood gases
(Peterson) 1982 First fibre optic-based biosensor for glucose 1983 First surface plasmon resonance (SPR)
immunosensor 1984 First mediated amperometric biosensor:
ferrocene used with glucose oxidase for glucose detection
History of Biosensors
![Page 8: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/8.jpg)
1987 Blood-glucose biosensor launched by MediSense ExacTech
1990 SPR based biosensor by Pharmacia BIACore
1992 Hand held blood biosensor by i-STAT
1996 Launching of Glucocard
1998 Blood glucose biosensor launch by LifeScan FastTake
1998 Roche Diagnostics by Merger of Roche and Boehringer mannheim
Current Quantom dots, nanoparicles, nanowire, nanotube, etc
History of Biosensors
![Page 9: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/9.jpg)
Components of a Biosensor
Detector
![Page 10: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/10.jpg)
BiosensBiosensoror
Analyte
Sample handling/preparation
Detection
Signal
Analysis
Response
![Page 11: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/11.jpg)
TARGET ANALYTEWhat do you want to detect?
Molecule Protein, DNA, Glucose, Vitamin, Sugar, metal ion
Protein Glucose
DNA
Bacteria
![Page 12: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/12.jpg)
Sample handlingHow to do deliver the analyte to the sensitive region?
•(Micro) fluidics•Concentration (increase/decrease)
•Filtration/selection
![Page 13: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/13.jpg)
RECOGNITION
How do you specifically recognize the analyte?
Antibody DNA
Complementary DNA
Antigen
Other:
enzyme/substrate
PNA/DNA or PNA/RNA
![Page 14: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/14.jpg)
Detection/Recognition
How do you specifically recognize the analyte?
Antibody Enzyme
Active site
Fab
Fc
Cell
Membrane receptors
Polymer/Hydrogel
Competitive binding
– enzyme/substrate;– antigen/antibody;– DNA/DNA;– DNA/transcription activator;– mRNA/DNA;– PNA/DNA or PNA/RNA;– microorganism/substrate;
![Page 15: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/15.jpg)
SIGNAL
How do you know there was a detection ?
Specific recognition?
DNA PROTEIN
Complementary
Common signaling principles
Optical (Fluoresence, Scanometric)
Electrical (Voltammetry, Potentiometry, conductivity)
Mass (QCM,Piezoelectric)
HIGH SENSITIVITY
HIGH SELECTIVITY
![Page 16: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/16.jpg)
Avoiding false signals
Specific recognition
Non specific signal
False specific recognition?
![Page 17: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/17.jpg)
Improving SIGNAL....
Signal LOW
SECONDARY SIGNAL
AMPLIFIER
Signal HIGH
Magnectic bead,
fluorecent dye, enzyme etc
AMPLIFICATION
![Page 18: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/18.jpg)
Improving SIGNAL....
PCRPCR
AMPLIFICATION
ELISA(Immunoblothin
g)
ELISA(Immunoblothin
g)
![Page 19: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/19.jpg)
DRAWBACK
COMPLICATED
EXPENSIVE
LABOR INTENSIVE PROCEDURE
TIME CONSUMING
NARROW TARGET
QUANTITATION
!!!
![Page 20: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/20.jpg)
Data Analysis
Response variable (R) vs time(t):Example of response variables:Refractive indexPotentialCurrentFrequencyMassPressureTemperature
t
R
![Page 21: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/21.jpg)
BaselineShould be stable when there is no binding
Quantifying NoiseRoot mean square (RMS) of a
sample of data points for a given time
Stable baseline Drift baseline
t t
Quantifying DriftShift in the baseline
(RMS) shown as response units per
time
![Page 22: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/22.jpg)
Common signal error sources
Inhomogenous sample
Bubbles/flow artifacts
Temperature
Electromagnetic interferance
Electronic unstability
Unstable chip/detection layer
![Page 23: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/23.jpg)
Improved sensitivityActive sensor
detects the analyte
Reference sensor
Coated with inert material does not detect the
analyte
Sample
R1 R2
Output signalR=R1-R2 or R=R1/R2
The reference is exposed to the same kind of
disturbances as the active sensor. These effects are
cancelled out by taking the difference between the two
sensors
t t t
R1 R2 R
![Page 24: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/24.jpg)
Signal interpretation
Visual (example pregnancy test)
Automatic (Software)
Manual (Research Biosensor)
![Page 25: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/25.jpg)
Kinetic evaluation
Binding / no binding
Affinity (Ka / Kd and k_on and k_off)
![Page 26: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/26.jpg)
1. LINEARITY Linearity of the sensor should be high forthe detection of high substrate concentration.2. SENSITIVITY Value of the electrode response per substrate concentration.3. SELECTIVITY Chemicals Interference must be
minimised for obtaining the correct result.4.RESPONSE TIME Time necessary for having 95% of the response.
Basic Characteristics of a Biosensor
![Page 27: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/27.jpg)
Example of biosensors
Pregnancy test
Detects the hCG protein in urine.
Glucose monitoring device (for diabetes patients)
Monitors the glucose level in the blood.
![Page 28: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/28.jpg)
Example of biosensorsExample of biosensors
Infectous disease biosensor from
RBS
Old time coal miners’ biosensor
![Page 29: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/29.jpg)
Research BiosensorsResearch Biosensors
Biacore Biosensor platform
![Page 30: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/30.jpg)
FluorescenceDNA MicroarraySPR Surface plasmon resonanceImpedance spectroscopySPM (Scanning probe microscopy, AFM, STM)QCM (Quartz crystal microbalance)SERS (Surface Enhanced Raman Spectroscopy)Electrochemical
Typical Sensing Techniquesfor Biosensors
![Page 31: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/31.jpg)
Types of Biosensors
1. Calorimetric Biosensor
2. Potentiometric Biosensor
3. Amperometric Biosensor
4. Optical Biosensor
5. Piezo-electric Biosensor
![Page 32: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/32.jpg)
Piezo-Electric Biosensors
The change in frequency is proportional to the mass of
absorbed material.
Piezo-electric devices use gold to detect the specific angle at which electron waves are emitted when the substance is exposed to laser light or crystals, such as quartz, which vibrate under the influence of an electric field.
![Page 33: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/33.jpg)
Electrochemical Biosensors
• For applied current: Movement of e- in redox reactions detected when a
potential is applied between two electrodes.
![Page 34: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/34.jpg)
Potentiometric Biosensor
For voltage: Change in distribution of charge is detected
using ion-selective electrodes, such as pH-meters.
![Page 35: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/35.jpg)
Optical Biosensors
•Colorimetric for color Measure change in light adsorption
•Photometric for light intensityPhoton output for a luminescent or
fluorescent process can be detected with photomultiplier tubes or
photodiode systems.
![Page 36: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/36.jpg)
Calorimetric Biosensors
If the enzyme catalyzed reaction is exothermic,
two thermistors may be used to measure the difference in resistance between reactant and product and,
hence, the analyte concentration.
![Page 37: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/37.jpg)
Electrochemical DNA Biosensor
Steps involved in electrochemical Steps involved in electrochemical DNA hybridization biosensors:DNA hybridization biosensors:
Formation of the DNA recognition layerFormation of the DNA recognition layer
Actual hybridization eventActual hybridization event
Transformation of the hybridization Transformation of the hybridization
event into an electrical signalevent into an electrical signal
![Page 38: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/38.jpg)
Motivated by the application to clinical diagnosis and genome
mutation detection
Types DNA Biosensors Electrodes
Chips
Crystals
DNA biosensor
![Page 39: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/39.jpg)
Wearable Biosensors
Ring Sensor
Smart Shirt
![Page 40: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/40.jpg)
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 anti- body based piezoelectric nanobiosensor to be used for anthrax,HIV hepatitis detection.
![Page 41: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/41.jpg)
Potential Applications
• Clinical diagnostics• Food and agricultural processes• Environmental (air, soil, and water) monitoring• Detection of warfare agents.
![Page 42: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/42.jpg)
Food Analysis Study of biomolecules and their interaction Drug Development Crime detection Medical diagnosis (both clinical and laboratory use)(both clinical and laboratory use) Environmental field monitoring Quality control Industrial Process Control Detection systems for biological warfare agents Manufacturing of pharmaceuticals and replacement organs
Application of Biosensor
![Page 43: MICROBIAL DETECTION USING BIOSENSOR. WHAT IS BIOSENSOR ? Analytical device which utilize biological reaction of biochemical molecule for detecting target](https://reader030.vdocument.in/reader030/viewer/2022012308/56649dff5503460f94ae789e/html5/thumbnails/43.jpg)