)2( ‫الكترونيات حيوية و قياسات‬

BBM322- Biomedical instrumentation II

Chapter 5: Biosensors
Prepared by:
Pr. Noman AL Naggar- Professor in biomedical engineering
Department of Biomedical Engineering UST-Sana’a
[email protected]
Modified by:
Dr. Nasr Kaid Alawdi- PhD in Biomedical Engineering,
Tlemcen University, Algeria.
Department of Biomedical Engineering UST-Aden
[email protected]
1
 Chapter 5: Biosensors

Outline ------------------------
1. Biosensors Definition
2. Components and Elements
3. Biosensing Principles
1. Enzymatic biosensors
2. Biosensor -Immunosensors
3. Biosensor -SPR biosensors
4. Biosensor - BIACORE biosensors
5. Biosensor -Fiber optic based biosensors
4. References

Slide-2
 Biosensor
What are biosensors?
• Biosensor: is a sensor using a living component or a product of a
living thing for measurement or indication. Or

A Biosensor Self-contained integrated device that is capable of

providing specific qualitative or semi-quantitative analytical
information using a biological recognition element which is in direct-
spatial contact with a transduction element. (IUPAC,1998)
Block diagram with components and according sensors types

4
COMPONENTS

DR.NOMAN AL NAGGAR ,USTY 5

ELEMENTS OF BIOSENSORS

DR.NOMAN AL NAGGAR ,USTY 6

 Biosensing Principles

 Analyte diffuses from the solution to the surface of the

Biosensor.
 Analyte reacts specifically & efficiently with the Biological
Component of the Biosensor.
 This reaction changes the physicochmical properties of the
Transducer surface.
 This leads to a change in the optical/electronic properties of
the Transducer Surface.
 The change in the optical/electronic properties is
measured/converted into electrical signal, which is detected.
 TYPICAL SENSING
TECHNIQUES
• Fluorescence.
• DNA Microarray.
• SPR (Surface Plasma Resistance).
• Impedance Spectroscopy.
• SPM (Scanning Probe Microscopy, AFM,
STM).
• QCM (Quartz Crystal Microbalance).
• SERS (Surface Enhanced Raman
Spectroscopy).
• Electrochemical.
 Nanosensor technology

LABEL-FREE LABEL

In labeled technology, some sort of label

has to be attached to the biomarker,
which otherwise would pass by
undetected…
 Labeled technology examples
labeled Non-
labeled
• Quantum
dots
• Gold
nanoparticles
• Radioactive
inks
 Why would I prefer a label-free
approach?

1. One fewer step to worry about (labeling)

2. I do not need a device to excite and image the

sample

3. I might create a lab-on-a-chip device

4. I would end-up with point of care (POC)

testing
APPLICATIONS
 Biosensor example in medical application
1. Enzymatic biosensors

Enzymes are catalysts for biochemical

reactions; large macromolecules
consisting mostly of protein, and usually
containing a prosthetic group (metals)
Substrate: the target molecule of an
enzymatic reaction

S: substrate; E: enzyme;
ES: enzyme-substrate complex;
P: product

DR.NOMAN AL NAGGAR ,USTY Slide-13

 Biosensor- 1. Enzymatic biosensors

• Advantages
– Highly selective
– Enzymes are catalytic, thus improving the sensitivity
– Fairly fast
• Disadvantages
– Expensive: cost of extraction, isolation and purification
– Activity loss when enzymes are immobilized
– Enzymes tend to be deactivated after a relatively short period of time
- In an enzymatic biosensor, the enzyme is immobilized as the
“receptor”
- Enzymes are specific to their substrates which can be the analyte
Example: glucose sensors
DR.NOMAN AL NAGGAR ,USTY Slide-14
 Biosensor- 1. Enzymatic biosensors
Enzymatic Approach (Glucose Sensors)

• Makes use of catalytic (enzymatic)

oxidation of glucose.
• The setup contains an enzyme electrode
and an oxygen electrode and the difference
in the readings indicates the glucose level.
• The enzyme electrode has glucose oxidase
immobilized on a membrane or a gel
matrix.

DR.NOMAN AL NAGGAR ,USTY Slide-15

 Biosensor- 1. Enzymatic biosensors
Affinity Approach -Glucose Sensors

• This approach is based on the

immobilized competitive binding of a
particular metabolite (glucose) and its
associated fluorescent label with
receptor sites specific to the
metabolite (glucose) and the labeled
ligand.
• This change in light intensity is then
picked up. Immobilized
Con A

DR.NOMAN AL NAGGAR ,USTY Slide-16

 Biosensor 1. Enzymatic biosensors
Alternatively, the reaction product hydrogen peroxide can also be
measured through reduction of H2O2

Use a Clark-type electrochemical cell similar to the one for measuring

O2, but at a positive bias voltage of 0.65V
Problems:
1. The reduction/oxidation potentials are fairly high ⇒ other materials
will interfere
2. The concentration of O2 needs to be kept constant, which is difficult
since O2 is involved in the reactions
DR.NOMAN AL NAGGAR ,USTY Slide-17
 Biosensor 2-Immunosensors
Immunosensors: based on highly specific antigen-antibody reactions
Antibody: works against the foreign body
Immunoglobin: globular proteins that participate in the immune response.
Structure antibody

The Fc end of an antibody can bind to other immune cells and the binding activate
these immune cells DR.NOMAN AL NAGGAR ,USTY Slide-18
 Biosensor 2-Immunosensors
• Detection of the reaction (binding/association)
can be either direct or indirect (labeled)
Direct detection: the binding of the analyte
(antigen) to the “receptor” (antibody) is detected
directly through the presence of the analyte

However, most are indirect due to lack of

signal from the analyte itself

• Indirect detection: signal is provided

by an exogenous reporter

DR.NOMAN AL NAGGAR ,USTY Slide-19

 Biosensor 2-Immunosensors

Indirect detection
• Radioimmunoassay (RIA)
– The labels are radioactive isotopes (for
example 131I)
– Highly sensitive and specific but
inconvenient
and expensive
– Less popular now due to safety issues
• ELISA (Enzyme-Linked
Immunosorbent Assay)
– Enzymes are used for labeling and
signal is provided by the reaction product
– Simple in terms of procedure and
equipment
• Fluorescent immunoassay
– Labeled with fluorescent dyes

Slide-20
Biosensor 2-Immunosensors

DR.NOMAN AL NAGGAR ,USTY Slide-21

 Biosensor 3-SPR biosensors
SPR (Surface Plasmon resonance)

Surface Plasmon's, are surface electromagnetic waves that propagate parallel along
a metal/dielectric interface
 Biosensor 3-SPR biosensors

• Direct detection – Surface Plasmon resonance.

• A surface plasma wave (SPW) is an electromagnetic wave (due to charge density
oscillation) which propagates along the boundary between a dielectric (e.g. glass)
and a metal.
• A surface plasmon wave can be excited optically: when the propagation
constant β of the incident light and the SPW are equal in magnitude and direction for
the wavelength.
• This corresponds to a certain angle of incident
light at a fixed wavelength ⇒ the reflected light
reaches its minimum.
• Binding induced change in propagation constant
of SPW is proportional to
the refractive index change in
the sample ⇒ the angle of
incident light also change (fig.)

DR.NOMAN AL NAGGAR ,USTY Slide-23

 Biosensor 3-SPR biosensors
Different parameters are measured in SPR biosensors to indicate the refractive index
change in the sample (due to binding)
Character of SPR biosensors:
• Direct detection ⇒ no need for labeling
• High sensitivity
• Fast ⇒ real-time monitoring
• Used with sample in liquid ⇒ important for biological samples
• Relatively simple device, which makes multichannel parallel detection easier ⇒high
throughput.
SPR commercialization - BIACORE
BIACORE optical system:
light from different angle of reflection is imaged onto
different position of a photo-detector array.

The corner-stones of Biacore technology

DR.NOMAN AL NAGGAR ,USTY Slide-24

 Biosensor 3-SPR biosensors
Surface Enhanced Raman Spectroscopy - SERS

•Nanostructured surface gives 10^6- q10^9 increase in signal

•High sensitivity, on par with heterogeneous, amplified systems
•SERS signal is photostable and, unlike fluorescence, cannot be “quenched”
•Signal is unaffected by ionic strength or temperature variations
•Does not work for all analytes
 Biosensor - 4. BIACORE technologhy biosensors

BIACORE features
• Specificity: different molecules
interact with a single partner
immobilized on a sensor surface
• Kinetics: rates of complex
formation (ka) and dissociation
(kd)
• Affinity: the level of binding at
equilibrium (KD, KA)
• Concentration: can be
determined for purified
molecules or for molecules in
complex mixtures such as serum
(needs a calibration curve)
• Multiple interactions during
Multiple flow-cells ⇒ simultaneous detection of reflection intensity
complex formation
Slide-26
 Biosensor – 5. Fiber optic based biosensors
• Optical fiber sensor – examples
• Sensing area along side of an optical fiber

• Sensing area located at distal end surface of an optical fiber

DR.NOMAN AL NAGGAR ,USTY Slide-27

 References
Medical Instrumentation: Application and Design, edited by John G. Webster
• Chemical sensors and biosensors, by Brian R. Eggins
• Sensors in Biomedical Applications: Fundamentals, Technology & Applications, by Gábor Harsányi
– Ch7: Biosensors
• Information about glucose meters
http://www.diabetesmonitor.com/meters.htm#fcnim
• Biomolecular sensors, edited by Electra Gizeli & Christopher R. Lowe
“Whole cell- and protein-based biosensors for the detection of bioavailable heavy metals
in environmental samples” by Philippe Corbisiera, Daniel van der Leliea, at al. ELSEVISER
Analytica Chimica Acta 387 (1999) 235-244.
2. “DNA BASED BIOSENSORS” By Zhai Iuni-Iui, Cui hong and Yang Yuifu, ELSEVISER
Biotechnology Advances, Vol. 15. No. 1. pp. 43-58.1997.
3. “BIOSENSORS” by Frieder Schelfer and Florian Schubert Akademie der Wissenscha fien,
Zentralinstitut fur Molekularbiologie, Robert-Riissle-Strasse lO,0-1115 Berlin-Buck
Germany ELSEVIER Amsterdam - London - New York -Tokyo 1992

DR.NOMAN AL NAGGAR ,USTY Slide-28