BE 247B: Topics Covered –Content/Schedule:

Topics Covered and Schedule:

1. (Jan 9) Introduction – What is Bioelectronics?
2. (Jan 11) Introduction to Biomaterials (structure
and function of proteins, nucleic acids, carbohydrates)
3.(Jan 16) Introduction to Biosensors (from single-use strips to
closed- loop systems.
4. (Jan 18) Protein (enzyme) bioelectronics
5. (Jan 23) Enzyme electrodes (from diabetes and kidney)
6. (Jan 25) Protein Immunoassays
7. (Jan 30) Exam 1
8. (Feb 1) DNA Bioelectronics
9. (Feb 6) DNA conductivity / Nanopores
10. (Feb. 8) Aptamer Bioelectronics
11.(Feb. 13) Wearable and Implantable Bioelectronic Systems
12. (Feb. 15) Wearable and Implantable Bioelectronic Systems (Part
2).
13. (Feb. 20) Flexible and Transient Bioelectronics
14. (Feb 22) Exam 2
15. (Feb 27) Closed loop (‘Sense-Act’) systems: Artificial Pancreas
16. (Feb 29) Closed loop systems: Beyond Diabetes (PD,
Anesthesia, Wound)
17. (March 5) a. Biofouling; b. Ingestible bioelectronic capsules
18. (March 7) Pacemakers, Brain Bioelectronics
19. (March 12) Bioenergy harvesting (BFC)
20. (March 14) Summary Future Prospects for Bioelectronics
21. (March 19) Final exam (3-6 pm)
An immunosensor is a biosensor in which a specific target
analyte, antigen, is captured by an immobilized antibody Detecting Antigen proteins using
followed by the formation of a stable immune complex and a Antibody Receptors
generation of an electrical signal proportional to the level of the
target antigen

Protein Function: Antibodies are Y-shaped proteins that bind

to the body's foreign invaders and signal the immune system to
get to work. These specialized, Y-shaped proteins that bind like
a lock-and-key to the body's foreign invaders — whether they
are bacteria, viruses or parasites.
Main components of an electrochemical immunosensor
 Electrochemical immunosensors (using enzyme tags, E) based on the
Sandwich or Competitive modes of operation.

Sandwich

Competitive

Competitive and Sandwich Immunoassays

Required Reading: Electrochemical immunosensors–a powerful tool for analytical

applications L Angnes - Biosensors and Bioelectronics, 102, 470 (2018).
Sandwich Immunoassays – use of Nanoparticle Tags
(coupled with ultrasensitive electrochemical stripping
detection of the Cd)

CdS CdS
32.00
c
u
2
22.75
C u r r e n t / A

u 1
b
i/A

13.50 0
0 30 60
u AFP concentration /
ppt a
4.25u
(CH2)1C0 ONH

(CH2)5OH
(CH2)5OH
(CH2)5OH
(CH2)5OH
(CH2)1C0

(CH2)1C0

-
ONH

-1.0 -0.9 -0.8 -0.6 -

NOH

5.00u
0.5
Square-wave voltamEm/oVgrams for a)
40 and c) 60 ppt AFP using quantum-dot
S S S S S 20, b) labeled AFP antibody
S S
Au

CdS Nanocrystal Tags for Protein

Detection: HIGH SENSITIVITY AND
SELECTIVITY
Label-free Bioelectronic Multiplexed Detection of Immunoassay: 1 D Nanomaterials (nanowires)

Massive redundancy in nanowire sensor

arrays.
NW sensor arrays with high
Nanowire-antibody density depending on the
ability to address the
hybrids for label-free individual nanowires.
biosensing Very dense arrays can be
fabricated.
BE/ECE/NE 247- Bioelectronics

Lecture 7 (Cont.)
DNA-BASED BIOELECTRONICS
 Schematic layout of a
Biosensor

Signal
E Transducer Signal
processor

Analyte • Optical
(substrate) • Electronic
• Thermometric
Biological recognition element • Piezoelectric
• Enzymes
• Microorganisms
• Antibodies (for ImmunoSensors)
• Tissues
• Nucleic acids (DNA,RNA) – Also Bioaffinity Assay (Like
Immunosensors
• aptamers
• Lectins
Biosensor
History
 Towards DNA Hybridization Sensors

DNA Hybridization Sensors

Detecting the order of the four chemical nucleobase building blocks

(i.e. the Sequence CCCACGGTCCCCAAG)

Transducer: This is the component that converts one form of energy into another. In the context of DNA hybridization sensors, the
transducer converts the biological reaction (DNA hybridization) into a measurable signal.

Signal: After the transduction process, the signal is often an electrical one that can be quantified and analyzed. This signal is
proportional to the concentration of the target DNA sequence in the sample.

Sample: This is the biological specimen containing DNA sequences that need to be analyzed. It could be from various sources, such
as blood, saliva, or environmental samples.

DNA Hybridization: This process involves the binding of a single-stranded DNA (ssDNA) probe to its complementary DNA
sequence in the sample. When the probe finds its complement, it forms a stable double-stranded DNA (dsDNA) hybrid.

Antibody (Ab): Antibodies can be used to recognize and bind to specific proteins or enzymes tagged to the DNA or its
complementary sequence. This binding event can facilitate the transduction of the hybridization event into a detectable signal.

Enzymatic (Enz.) Reaction: An enzyme linked to the detection process catalyzes a reaction that produces a detectable change, often
resulting in a colorimetric, fluorescent, or electrochemical signal.

Immunological Reaction: This likely refers to the use of an immunological event, such as the specific binding of an antibody to a
target, which can be part of the detection mechanism.

Detecting the Order of Nucleobase Building Blocks: The system is capable of identifying the specific order of the nucleobases in a
DNA strand, which is crucial for identifying genetic sequences. The example given, "CCCACGGTCCCCAAG," is a sequence of 15
nucleobases.
Intelligent Questions and Answers:
Q1: How does the DNA hybridization sensor differentiate
between similar DNA sequences?
A1: The DNA hybridization sensor relies on the principle of
complementary base pairing, where the ssDNA probe will only form a
stable hybrid with a perfectly complementary sequence. Mismatches
in the base pairing can result in weaker binding or no binding at all,
which can be detected by the transducer as a change (or lack of
change) in the signal.

Q2: What types of signals can a transducer produce in a DNA

hybridization sensor system?
A2: A transducer can produce various types of signals, including
electrical, optical (such as fluorescence or color change), or thermal.
The type of signal depends on the design of the sensor and the
detection method employed.

Q3: Why might an enzyme be used in conjunction with a DNA

hybridization sensor?
A3: Enzymes can amplify the signal from the hybridization event,
making it easier to detect. For instance, an enzyme might catalyze a
reaction that generates a large number of colored or fluorescent
molecules for each hybridization event, significantly increasing the
signal's intensity.

Q4: What are the potential limitations of using antibodies in a

DNA hybridization sensor system?
A4: Antibodies may have cross-reactivity, which can lead to non-
specific binding and false positives. They also require careful storage
and handling as they are sensitive to environmental conditions.
Additionally, the production of specific antibodies can be time-
consuming and costly.

Q5: In what way can the immunological reaction contribute to the

detection of DNA sequences?
A5: Immunological reactions can be used to produce a measurable
response to the presence of the target DNA sequence. For example,
if the target DNA or the probe is tagged with an antigen, the
corresponding antibody can bind to it, and this event can be coupled
to a signal-generating system.

Understanding the principles behind each component and how they

work together is crucial for mastering the operation of DNA
hybridization sensors.
Why DNA / RNA?
DNA is the genetic code of life, the instructions for building
and operating an organism. RNA is primarily a messenger
molecule, carrying instructions from the DNA code to control
the synthesis of proteins — the building blocks of organisms.
While DNA contains the sugar deoxyribose
RNA contains the sugar ribose.

DNA sequencing can answer a range of biological questions,

providing information on pathogen identity, genetic disease
risk or how an organism has evolved. Sequencing DNA means
determining the order of the four chemical building blocks -
called "bases" - that make up the DNA molecule. Sequence
data can highlight changes in a gene that may cause disease.
While DNA contains the
sugar deoxyribose RNA
contains the sugar ribose.

The text outlines the roles of DNA and RNA in living organisms. DNA is described as
the "genetic code of life," containing instructions for building and operating an
organism. RNA is primarily a messenger molecule that carries instructions from the
DNA to control protein synthesis, which forms the building blocks of organisms. The
text also highlights the difference in the sugar components of DNA and RNA: DNA
contains deoxyribose, while RNA contains ribose.

DNA sequencing is presented as a valuable tool for answering biological questions,

identifying pathogens, assessing genetic disease risk, and understanding evolutionary
processes. Sequencing DNA involves determining the order of the four chemical bases
(adenine, thymine, guanine, and cytosine) that make up the DNA molecule. The
sequence data can reveal changes in genes that may lead to disease.

Molecular Diagram Explanation:

The diagram shows a section of a DNA molecule, highlighting its double-stranded
structure with base pairs connected between the two strands. The sugar-phosphate
backbone is illustrated, with the sugars represented in pink (deoxyribose) and the
phosphate groups in yellow. The four bases (adenine, thymine, guanine, and cytosine)
are shown with their standard color coding: adenine in blue, thymine in red, guanine in
green, and cytosine in yellow. Hydrogen bonds between the bases are shown as dotted
lines, indicating base pairing (adenine with thymine, and guanine with cytosine).
Intelligent Questions and Answers:

Q1: Why is the sugar component different in DNA and RNA, and what implications does
this have for their function?
A1: The sugar component differs in DNA (deoxyribose) and RNA (ribose) because of the
presence of one less oxygen atom in DNA. This small structural difference makes DNA
more chemically stable and less reactive than RNA, which is suitable for the long-term
storage of genetic information. RNA, being more reactive, is suitable for tasks that
require a molecule that can easily be synthesized and degraded, such as coding,
decoding, regulation, and expression of genes.

Q2: What is the significance of the specific base pairing in DNA?

A2: The specific base pairing (adenine with thymine, and guanine with cytosine) is
critical for the accurate replication of DNA during cell division. It ensures that the genetic
information is copied correctly, allowing for the precise transmission of genetic
instructions from one generation to the next.

Q3: How can changes in the DNA sequence lead to disease?

A3: Changes or mutations in the DNA sequence can alter the structure and function of
the proteins produced. If these proteins play critical roles in cellular processes, their
malfunction can lead to disease. For example, a single base change in the gene
responsible for hemoglobin can lead to sickle cell anemia, a condition where the altered
hemoglobin causes red blood cells to become rigid and sickle-shaped.

Q4: In what ways has DNA sequencing impacted our understanding of human evolution?
A4: DNA sequencing has allowed scientists to compare the genomes of different
species, identifying genetic similarities and differences. This has provided insights into
the evolutionary relationships between species, the identification of evolutionary
conserved sequences important for life, and the understanding of how certain traits have
evolved.

Q5: What are some of the challenges of DNA sequencing technology?

A5: DNA sequencing technology faces challenges such as high costs for whole-genome
sequencing, the need for high-quality DNA samples, and the complex data analysis
required to interpret the vast amount of data generated. Additionally, ethical issues arise
regarding privacy and the use of genetic information.
 DNA Hybridization

DNA molecule consists of two coiled strands run

in antiparallel direction, and held together by
Hybridization is the formation of the double helix (ds hydrogen bonding between complementary
DNA) by base pairing between complementary strands. bases.
DNA hybridization is the process of combining two complementary single-stranded DNA molecules
and allowing them to form a single double-stranded molecule through base pairing.

Probe
DNA biosensors consist of an
immobilized DNA strand to
detect the complimentary
sequence (target DNA) by
DNA–DNA hybridization.
The probes are typically
short oligonucleotides (25 -
40 mer) that are capable of
hybridizing with specific and
unique regions of the target
nucleotide sequence.
Target
Three key steps :
G (i) probe immobilization
G =C
A A = T on a solid substrate, (ii)
hybridization (or
T T =A
G recognition) of the
G =C complementary target
T Hybridization Signal
T =A strand and (iii)
A A = T conversion of the
C C =G hybridization event into
C C =G a physical signal
T T =A (i.e. transduction).
G G =C

DNA HYBRIDIZATION BIOSENSORS

DNA Hybridization Sensors
Are based on: DNA Structure – Base Pairing
(between single strands)

Only certain pair bases can form these bonds.

Adenine pairs with thymine (via 2 hydrogen bonds)
while guanine pairs with cytosine (via 3 such bonds).
Such paired DNA strands are said to be
complementary.
The genetic information is embodied in the sequence of
the four bases (e.g., GGADTTTACAACC).

Hydrogen bonding between complementary

bases.
 Why DNA Biosensors?
APPLICATIONS OF DNA
HYBRIDIZATION
• BIOSENSORS
SCREENING FOR GENETIC OR INFECTIOUS DISEASES

• ON-SITE ENVIRONMENTAL DETECTION OF VIRAL OR

BACTERIAL PATHOGENS

• FORENSIC INVESTIGATIONS IN CRIME SCENES

• DETECTION OF FOOD CONTAMINANTS

• DETECTION OF BIOLOGICAL WARWARE AGENTS

Hybridization is the basis of DNA biosensors.

Immobilized ssDNA probe

Steps involved in the detection of a DNA

sequence.
 DNA HYBRIDIZATION BIOSENSORS

How to Detect the Hybridization Event?

Three key steps have to be

controlled in a DNA biochip: (i)
probe immobilization on a solid
substrate, (ii) hybridization (or
recognition) of the complementary
target strand and
(iii) conversion of the
hybridization event into a
physical signal
(i.e., transduction).

Good Review:
DNA electrochemistry and electrochemical sensors for nucleic
acids; Annual Review of Analytical Chemistry, 11, 197(2018 )
Surface Chemistry:
The immobilization of the nucleic acid probe onto the
transducer surface plays an important role in the
overall performance of DNA biosensors and gene chips.

The immobilization step should lead to a well-defined probe

orientation, readily accessible to the target. Depending upon the nature
of the physical transducer, various schemes can be used for attaching
the DNA probe to the surface.

These include thiolated DNA for self assembly onto gold transducers
the use of (gold electrodes or gold-coated piezoelectric crystals),
covalent linkage to the gold surface via functional alkanethiol-based
monolayers, the use of biotylated DNA for complex formation with a
surface-confined avidin or strepavidin, covalent (carbodiimide)
coupling to functional groups on carbon electrodes, or a simple
adsorption onto carbon surfaces. As in solution-based hybridization
assays, conditions for interfacial hybridization events (e.g., ionic
strength, temperature, presence of accelerators) have to be optimized.
Chemical and thermally-induced dehybridization of the resulting
duplex is often used for regenerating the interface.
 Surface
Immobilization
The probe immobilization step plays a major role in the
overall performance of DNA biosensors. The achievement
of high sensitivity and selectivity requires maximization
of the hybridization efficiency and minimization of non-
specific adsorption events, respectively. The probes are
typically short oligonucleotides (25 - 40 mer) that are
capable of hybridizing with specific and unique regions of
the target nucleotide sequence.

Control of the surface chemistry and coverage is essential

for assuring high reactivity, orientation/accessibility,
and stability of the surface-bound probe, as well as for
avoiding non-specific binding/adsorption events.
 DNA Bioelectronics (probe immobiliz.)
(towards DNA Hybridization)

Thiolated DNA for self assembly onto gold electrode

transducers

Alkanethiol self-assembled
monolayers (SAMs) have seen
widespread utility in the fabrication
of electrochemical biosensors,
e.g., immobilizing DNA probes
(ssDNA).

The affinity of thiols for

the gold surface makes
alkanethiols ideal for the
preparation of modified
electrodes.

Immobilizing the ssDNA receptor with gold electrode

transducer
 Use of mixed SAM layers for simultaneous DNA orientation
control and surface blocking (against non-specific adsorption)

After treatment with mercaptohexanol (MCH), a short

alkanethiol with a terminal hydroxy group, the DNA probe
“stands up” and extends farther into the solvent phase.
 Surface
Immobilization
The probe immobilization step plays a major role in the
overall performance of DNA biosensors. The achievement
of high sensitivity and selectivity requires maximization
of the hybridization efficiency and minimization of non-
specific adsorption events, respectively. The probes are
typically short oligonucleotides (25 - 40 mer) that are
capable of hybridizing with specific and unique regions of
the target nucleotide sequence.

Control of the surface chemistry and coverage is essential

for assuring high reactivity, orientation/accessibility,
and stability of the surface-bound probe, as well as for
avoiding non-specific binding/adsorption events.
 Immobilization of the ssDNA Probe - Alkanethiol self-assembly

The probe immobilization step plays a Mixed monolayers of thiol-derivatized single-stranded

major role in the overall performance oligonucloetide and 6-mercapto-1-hexanol (MCH) to
of DNA biosensors. minimize non-specific adsorption

The achievement of high sensitivity and

selectivity requires maximization of the
hybridization efficiency and
minimization of non-specific adsorption
events, respectively.

Control of the surface chemistry and

coverage is essential for assuring high
reactivity, orientation/accessibility,
and stability of the surface-bound
probe, as well as for avoiding non-
specific binding/adsorption events.
 Surface Immobilization (Cont.)
Common probe immobilization schemes include attachment of biotin-functionalized probes to
avidin-coated surfaces, self-assembly of organized monolayers of thiol functionalized probes
onto gold transducer, carbodiimide covalent binding to an activated surface,
electropolymerization of conducting polymers, as well as adsorptive accumulation onto
carbon- paste or thick-film carbon electrodes.

The use of alkanethiol self-assembly methods have been particularly attractive for
fabricating reproducible probe-modified surfaces with high hybridization activity. For this
purpose, the DNA is commonly immobilized on gold by forming mixed monolayers of
thiol- derivatized single-stranded oligonucloetide and 6-mercapto-1-hexanol.
 Alkanethiol self-assembly
Mixed monolayers of thiol-derivatized single-stranded
oligonucloetide and 6-mercapto-1-hexanol (MCH) to
minimize non-specific adsorption
Bioelectronic DNA Hybridization Sensors: These can be divided into two
major principles, involving the
How to Detect the Hybridization Event? use of labels generating an
electrical signal or label-free
Bioelectronic signal transduction through: protocols.

• REDOX INDICATORS
(INTERCALATORS)
• LABEL-FREE DETECTION: INTRINSIC ELECTROACTIVITY
OF DNA
• ENZYME TAGS
• CHANGES IN INTERFACIAL PROPERTIES (e.g.,
Impedance)
• METAL NANOPARTICLE TAGS
SIGNAL-GENERATING MECHANISMS
 ELECTRICAL
TRANSDUCTION:
SIGNAL-GENERATING MECHANISMS

• REDOX INDICATORS
(INTERCALATORS)
• LABEL-FREE DETECTION:
INTRINSIC
ELECTROACTIVITY OF
DNA
• ENZYME TAGS
• CHANGES IN
INTERFACIAL
PROPERTIES
• METAL NANOPARTICLE
TAGS
 Redox Hybridization Indicators
Redox indicators are small electroactive
DNA-intercalating or groove-binding
substances, that posses a much higher
affinity for the resulting ds hybrid
compared to the ss single-stranded probe.
Accordingly, the concentration of the
indicator at the electrode surface increases
when hybridization occurs, resulting in
increased electrochemical response.
 USE OF REDOX INDICATORS (INTERCALATORS)

Electroactive indicators for monitoring the hybridization event:

Detecting the hybridization event via the increased current signal of

an electroactive indicator (that preferentially binds to the DNA
duplex).

Such redox-active compounds have a much larger affinity for the

resulting duplex (compared to their affinity to the probe alone).

Their association with the surface duplex thus results in an increased

electrochemical response.
 REDOX HYBRIDIZATION INDICATORS

Threading intercalator ferrocenyl naphthalene diimide binds to the

DNA duplex more tightly than usual intercalators (b) and displays
a negligible affinity to the single-stranded probe (a). Adding
ferrocene redox group to the aromatic (intercalating) structure.
(Takenaka et al, AC 2000)
 Electrical signal transduction of DNA Hybridization
through: Use of REDOX INDICATORS (INTERCALATORS)

intercalation is the insertion of a molecule (or a group of molecules) between two other
molecules (or groups). In the present approach, a Redox molecule or a complex of Redox
molecules are employed for DNA strand binding via intercalation. The binding to ssDNA and
dsDNA is obtained with different affinity regarding the nature of the intercalator.

This includes organic molecules such as the commonly used MB as well as oracet blue
(OB) and toluidine blue. Various other molecules and macromolecules could also be
intercalated as an organometallic complex including Ru(NH3)63+ , cobalt phenanthroline, or
metal intercalating agents such as palladium nanoparticles
 ELECTRICAL
TRANSDUCTION:
SIGNAL-GENERATING MECHANISMS

• REDOX INDICATORS
(INTERCALATORS)
• LABEL-FREE DETECTION:
INTRINSIC
ELECTROACTIVITY OF
DNA
• ENZYME TAGS
• CHANGES IN
INTERFACIAL
PROPERTIES
• METAL NANOPARTICLE
TAGS
ENZYME TRACERS

The use of enzyme tags to generate

electrical signals offers great promise for
ultrasensitive electrical detection of
DNA hybridization (in a manner
analogous to their use in
immunosensors).

Commonly, enzymes such as alkaline

phosphatase (ALP), peroxidase (HRP) or
glucose oxidase, generating electroactive
species.

(Read: Electroanalysis 9(1997)737)

Use of ENZYME TAGS-
Sandwich Assay:
Steps involved in the bioelectronic DNA detection: A) Hybridization; B)
Amperometric detection. Capturing an E-tag sequence following by adding
its substrate S of E to detect the reaction product P.
Using horseradish-peroxidase (HRP) Enzyme Tag
 Enzyme (HRP) TAG – Mismatch Discrimination

Highly sensitive and selective amperometric monitoring of DNA hybridization based on

the use of horseradish-peroxidase (HRP) labeled target and an electron-conducting redox
Os polymer. Current increments upon raising the hydrogen peroxide concentration from 0
to 1 mM. (a) Without the analyzed sequence in the droplet; (b) with 0.1 fM perfectly
matched analyzed sequence; (c) as in (b), but with a mismatched base; (d) as in (b), with
two mismatched bases.
 Towards DNA Arrays:

Ultimately, these developments

will lead to the introduction of
miniaturized (on-chip) sensor
arrays, containing numerous
microelectrodes (each coated with
a different oligonucleotide probe)
for the simultaneous hybridization
detection of multiple DNA
sequences.
 GeneFluidics 16 sensor DNA
array for pathogen
identification

GeneFluidics electrochemical biosensor composed of an

array of 16 sensors. The sensors have been functionalized
with pathogen-specific DNA capture probes in duplicates. EC=E. coli;
PM=Proteus mirabilis, KE=Klebsiella-Enterobacter,PA=Pseudomonas aeruginosa,
EF=Enterococcus, UNI=universal bacterial probe, EB=Enterobacteriaceae, NC=
negative control (no target).
GeneFluidics electrochemical biosensor composed of an array of 16 sensors. The sensors have been functionalized with
pathogen-specific DNA capture probes in duplicates. EC=E. coli; PM=Proteus mirabilis, KE=Klebsiella-
Enterobacter,PA=Pseudomonas aeruginosa, EF=Enterococcus, UNI=universal bacterial probe, EB=Enterobacteriaceae,
NC= negative control (no target).
 DNA Chips:
Multiplexed Detection of Multiple DNA
Target

Detection of 64 multiple DNA sequences.

High density DNA array: Based on multiple ssDNA-

functionalized electrode transducers.
Bioelectronic DNA Hybridization Sensors:

How to Detect the Hybridization Event?

Electrical signal transduction through:

• REDOX INDICATORS (INTERCALATORS)

• LABEL-FREE DETECTION: INTRINSIC ELECTROACTIVITY
OF DNA
• ENZYME TAGS
• CHANGES IN INTERFACIAL PROPERTIES (e.g.,
Impedance)-Label-Free
• METAL NANOPARTICLE TAGS
SIGNAL-GENERATING MECHANISMS
 Another Label-Free DNA Detection:
Hybridization-induced changes in electrochemical
parameters (e.g., capacitance or conductivity).

Increased attention has been given recently to new indicator-free electrochemical

detection schemes that greatly simplify the sensing protocol.

Such direct, label-free, electrical detection of DNA hybridization can be

accomplished by monitoring changes in the conductivity of conducting
polymer molecular interfaces or 1D nanowires transducers.
Bioelectronic DNA Hybridization Sensors:

How to Detect the Hybridization Event?

Electrical signal transduction through:

• LABEL-FREE DETECTION: INTRINSIC ELECTROACTIVITY

OF DNA
• ENZYME TAGS
• CHANGES IN INTERFACIAL PROPERTIES (e.g.,
Impedance)
• METAL NANOPARTICLE TAGS

SIGNAL-GENERATING
MECHANISMS
 ELECTROCHEMISTRY OF DNA
Label-Free Detection based on the intrinsic electroactivity
of
DNA

REDUCTION SITE
Among the four nucleic acids bases, the guanine moiety
OXIDATION is most easily oxidized and is most
SITE

suitable for such indicator-free hybridization detection.

Electrochemistry of nucleic acids
E Palecek, M Bartosik - Chemical Reviews, 112, 3427 2012

DNA Electrochemistry

Carbon electrodes are probably the most frequently employed electrodes in studies
of oxidation of NA bases and their nucleotides or nucleosides. Purine bases
require lower overpotential for oxidation than pyrimidines, with G being the most
easily oxidized. Both A and G produce oxidation peaks in a wide pH range (0–12.5).
 ELECTRICAL
TRANSDUCTION:
SIGNAL-GENERATING
• REDOX MECHANISMS
INDICATORS
(INTERCALATORS)
• LABEL-FREE DETECTION:
INTRINSIC ELECTROACTIVITY OF
DNA
• ENZYME TAGS
• CHANGES IN INTERFACIAL
PROPERTIES
• METAL NANOPARTICLE TAGS
METAL NANOPARTICLE TAGS

ELECTROCHEMICAL CODING OF DNA

USE OF QUANTUM DOTS FOR MULTI-TARGET DETECTION

ZnS CdS PbS T1 T2 T3

P’1 P’2 P’3

T1 T2 T3

P1 P2 P3

JACS125(2003)3214

NANOPARTICLE-BASED ELECTRICAL DNA ASSAYS

(in connection to voltammetric detection of the
crystals)
 ENCODED BEADS FOR ELECTROCHEMICAL
IDENTIFICATION – TOWARDS PARTICLE-BASED LIBRARY
FOR ELECTRICAL CODING

3:1:1

T3
MB
T3
Current ( µA )

1:3:3

MB T2
T2
1:1:3

MB T1

T1 -1.0 -0.5
Potential ( V )

vs. ‘supermarket’ barcoding

 Nanoparticle-based Conductivity Detection of DNA Hybridization

A A Ag+ A

hydroquinone e-

Oligonucloetide probe immobilized in the gap between the two

microelectrodes. The hybridization event thus localizes gold
nanoparticles in the electrode gap, and along with subsequent silver
deposition leads to measurable conductivity signals. Such hybridization-
induced conductivity signals, associated with resistance changes across
the electrode gap, offer high sensitivity with a 0.5 picomolar detection
limit.
 DNA BIOSENSORS (Beyond Hybridization)

BASED ON DIFFERENT DNA BINDING PROCESSES

A B C D E
A. Hybridization to form Watson-Crick pair
B. Hoogsteen triplet formulation
C. Nucleic acid-protein interaction
D.Interaction of nucleic acid-protein complex
with small molecules and drugs (E)
Detecting DNA damage
Another active field is the integration of the
sample preparation and DNA array detection in
the s o - called ‘Lab-o n - a - Chip’ (LOC)
configuration.

The goal of this technology is to fully

integrate multiple processes, including
sample collection and pretreatment with the
DNA extraction, amplification, hybridization
and detection, on a microfluidic platform.

The ability to perform all the steps of the

biological assay on a single s e l f - contained
microchip promises significant advantages in
terms of speed, cost, sample/reagent
consumption, contamination, efficiency and
automation (including parallel sample
processing).
 GeneFluidics 16 sensor DNA
array for pathogen
identification

GeneFluidics electrochemical biosensor composed of an

array of 16 sensors. The sensors have been functionalized
with pathogen-specific DNA capture probes in duplicates. EC=E. coli;
PM=Proteus mirabilis, KE=Klebsiella-Enterobacter,PA=Pseudomonas aeruginosa,
EF=Enterococcus, UNI=universal bacterial probe, EB=Enterobacteriaceae, NC=
negative control (no target).