BIOSENSOR

BIOSENSOR
A biosensor is an analytical device, used for the detection of an analyte that combines a
biological component with a physicochemical detector.

Biosensors are diagnostic instrumentations that provide accurate on-the-spot results of

biochemical tests.

These instruments are designed based on a combination of principles from various areas—
biochemistry, membrane technology, and microelectronics.

A biosensor is more than a biological probe like a pH meter that measures a biochemical or
biological reaction. Integrated into the design and construction of a biosensor is a
biochemical component that allows signals from specific biochemical reactions/ physio-
iochemical change to be detected, measured, and documented.
 IUPAC CONCEPT
According to a recently proposed IUPAC definition, A biosensor is a self-contained integrated
device which is capable of providing specific quantitative or semi-quantitative analytical
information using a biological recognition element (biochemical receptor) which is in direct
spatial contact with a transducer element.

A biosensor should be clearly distinguished from a bioanalytical system, which requires

additional processing steps, such as reagent addition.

Furthermore, a biosensor should be distinguished from a bio-probe which is either disposable

after one measurement, i.e. single use, or unable to continuously monitor the analyte
concentration.
 WORKING PRINCIPLE
Three major components:
 Biological sensitive element
 Transducer
 Detector

The biologically sensitized elements (probe) include receptors, enzymes, antibodies, nucleic acids, molecular imprints, lectins,
tissue, microorganisms, organelles etc., which are either a biologically derived material or bio-mimic component that receives
signals from the analytes (sample) of interest and transmits it to transducer. Sometime, the physical change(allosteric) is the
only out put from the biocomponent and analyte interaction.
A transducer is basically a device that converts one form of energy to another. A biosensor utilizes a physiochemical
transducer that is in intimate contact with the biorecognition layer, essentially forming a biotransducer to convert chemical
energy into electrical energy. The transducer acts as an interface, measuring the physical or chemical change that occurs
during reaction between bio-receptor/sensitive biological element and analyte, then transforming that energy into measurable
electrical output. The intensity of generated signal is directly or inversely proportional to the analyte concentration.

The detector element traps the signals from the transducer, which are then passed to a microprocessor where they are
amplified and analyzed; the data is then transferred to user friendly output and displayed/stored

Difference between transducer and bio transducer????

 MEMBRANES USED IN BIOSENSORS

Substrate screening-protect larger size substrate from entrance

Screening of products-protect transducer from reacting with unwanted products

A membrane also limits the rate of diffusion of an analyte to the biological element of a
biosensor, thereby preventing high concentrations of the analyte from overwhelming the
sensor.

Such restrictions allow the instrument to achieve a linear response to the analyte
concentration
 CHARACTERISTICS OF AN IDEAL BIOSENSOR

Miniaturization of technology (implantable biosensors)

Highly specific, no false positive result should come
Reactions should be independent of environmental conditions

Response should be fast, accurate, specific and linear according to concentration of analytes
Reactions should be sensitive to small amount of analytes
In case of implantable biosensor it should be non toxic.-insulin pump

It should be cost effective

Structure should be robust and stable under harsh conditions
Half life of sensing material should be cost effective
 TERMINOLOGIES

Biological oxygen demand is the amount of dissolved oxygen needed by

aerobic biological organisms to break down organic material present in a given
water sample at certain temperature over a specific time period. Its a measure
of organic pollutants.

Chemical Oxygen Demand or COD(xenobiotics) is a measurement of

the oxygen required to oxidize soluble and particulate organic and inorganic
matter in water. The chemical oxygen demand (COD) test is commonly used
to indirectly measure the amount of pollutants present in water.

Why COD always has a higher value than BOD?

 PRINCIPLES OF MFC TECHNOLOGY

Microbial fuel cells are devices that directly convert the chemical energy in organic matter into electricity via metabolic
processes of microorganisms. An MFC comprises of two electrodes, an anode and a cathode, in the presence of an electrolyte.
The two electrodes are usually divided by a proton exchange membrane (PEM), and are connected by an external circuit that
includes an external load. Electroactive bacteria (anodophiles) reside at the anode of the device in the form of a biofilm.
The anodophiles oxidize the biodegradable organic molecules present in the feed solution and generate electrons, protons and
carbon dioxide. In the absence of oxygen, the electrons are extracellularly transferred to the anode and flow through the
external circuit towards the cathode thus producing electricity. Protons migrate through the PEM to the cathode and react
with electrons and an electron accepter (usually oxygen) to form water. The anode material must be porous and have a large
surface area to accommodate biofilm growth. The cathode is usually doped with catalysts, such as platinum, in order to
increase the rate of oxygen reduction reactions at the electrode surface.

The anodophiles in MFCs can degrade a multitude of organic molecules in wastewater, such as acetate, propionate, butyrate,
while simultaneously generating electricity. The most intuitive use of the MFC technology regards therefore the
development of devices that treat wastewater whilst generating electricity.
The current generated by an MFC directly proportional to the metabolic activity of the electroactive biofilm at the anode surface.
That means it indicates the amount of organic matters present in the sample
The anodic biofilm of the MFC acts as the recognition component (bioreceptor). Its response to the specific disturbance
affects the rate of flow of electrons to the anode (the transducer) and it is transduced into a measurable current change.
BOD is measured by calibration.
 ELECTROACTIVE BACTERIA AND THEIR
MECHANISM OF ACTION
In general, electroactive bacteria (EAB) are defined as being capable of transporting electrons over biological
membranes to or from their extracellular environment.

 Carrier in
membrane and out
side the membrane

 Carriers in pili
ENVIRONMENTAL APPLICATIONS OF BIOSENSOR
BOD5 Detection:

Oxygen can be provided by photosynthetic system

Ultimate Waste Water Treatment with corresponding electricity generation
biotransducer
Pesticide detection:
 Acetylcholine esterase enzyme based or antibody based biosensors are used for organophosphates detection.

Gas detection:
Species Gas
Methylomonas Methane
Pseudomonas Carbondioxide
Thiobacillus Sulphur dioxide
 OTHERS
Toxins and poison detection in usable water and sewage
Flow monitor device in waste water treatment plant

Pathogen detection
Nitrogen enriched compound detection to prevent eutrophication
Heavy metal detection
Poisonous gas and bio-weapon detection

Antibiotic detection for maintaining the ecological balance

Insecticides and herbicides detection
 ENRICHMENT TECHNIQUE
Enrichment media promote the growth of a particular microorganism against the others present in the sample by its specific
nutrient utilization ability or other unique metabolic properties (e.g. resistance to specific chemical). Only those microbes can
grow in the enrichment medium that can use or tolerate the components of the selective medium.

Sample Enrichment medium is designed in a way that

only organisms that can utilize the substrate as
carbon source will grow

Sample preparation
Organism
inoculation into enrichment medium
OD for growth check

Spread plate after desired serial dilution

Single colony isolation

Biochemical and
molecular
Streak plate for pure culture Desired assays characterization
(16s and 18s)
Lipid Degrading Bacteria