Biosensors

1
Biosensors

• Christopher Byrd
• ENPM808B
• University of Maryland, College Park
• December 4, 2007

2
Outline

• Introduction
• 4 Specific Types of Biosensors
• Electrochemical (DNA)
• Carbon nanotube
• BioFET
• Whole Cell
• Basic functionality
• Benefits/Challenges
• Summary
• References

3
Introduction

• Biosensor
• Incorporation of a biomolecule in order to detect
  something

Species to be detected (analyte)
Filter
Recognition Layer
Transducer
Electronics
Signal
Recognition Layer
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
4
Introduction

• Biosensors 3B
• 90 ? Glucose testing
• 8 - 10 increase in industry per year

Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
5
Electrochemical DNA Sensors

• Harnesses specificity of DNA
• Simple assembly
• Customizable
• Vast uses for small cost

Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
6
DNA Structure

• DNA structures---double helix
• 4 complementary bases
• Adenine (A), Guanine (G),
• Thymine (T), and Cytosine (C)

Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
7
DNA Specificity

• Hydrogen bonding between base pairs
• Stacking interaction between bases along axis of
  double-helix
• Animation

Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
8
Principles of DNA biosensors

• Nucleic acid hybridization

(Target Sequence)
(Hybridization)
(Stable dsDNA)
ssDNA (Probe)
Source http//cswww.essex.ac.uk
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
9
E-DNA Sensor Structure
Stem-loop
s
Gold electrode
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
10
E-DNA Sensor Structure
Target
Stem-loop
s
Gold electrode
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
11
E-DNA Sensor Structure
(Open, extended)
(Stem-loop)
Source Ricci et al., Langmuir, 2007, 23,
6827-6834
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
12
Carbon Nanotube Biosensor
Image www.cnano-rhone-alpes.org
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
13
Carbon Nanotube Biosensor

• One atom thick
• One nanometer diameter
• Ability to be functionalized
• Electrical conductivity as high as copper,
  thermal conductivity as high as diamond

Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
14
CNT Biosensor Structure
Succinimidyl ester
Source Chen et al., 2001
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
15
CNT Uncoated vs. Coated
Source Chen et al., 2001
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
16
Carbon Nanotube Biosensors
Source Chen et al., 2001
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
17
CNT Biosensor Signal Detection
Glucose
O2
Gluconic Acid
H2O2
e-
Source Besteman et al., 2003
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
18
CNT Biosensor Signal Detection
e-
e-
e-
e-
e-
Effectively increases electrical current
Source Besteman et al., 2003
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
19
CNT Biosensor Results
160 mM
60 mM
20 mM
0 mM
Source Besteman et al., 2003
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
20
BioFET

• Draws upon versatility of common electronic
  component (Field-Effect Transistor)
• Well understood expectations/results

Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
21
FET
-
Drain
Gate
Insulator
Source







Source Hayes Horowitz, 1989
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
22
FET

-
Drain
Gate
Insulator
Source




(Not conductive enough)
(Electron Channel)
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-
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
23
FET

-
Threshold Voltage
Drain
Gate
Insulator
Source




Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
24
FET

-
Drain
Gate
Insulator
Source








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Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
25
BioFET
Source Im et al., 2007
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
26
BioFET
Source Im et al., 2007
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
27
BioFET Results
Gate (before)
Source Im et al., 2007
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
28
BioFET Results
Gate (w/ complete Biomolecule)
Gate (after etch, w/biotin)
Source Im et al., 2007
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
29
Whole Cell Sensors
Source http//www.whatsnextnetwork.com/technology
/media/cell_adhesion.jpg
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
30
Whole Cell Sensors

• Harness normal genetic processes
• May detect dozens of pathogens
• Modifiable/customizable
• Reports bioavailability
• Temperature/pH sensitive
• Short shelf-life

Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
31
Whole Cell Sensors
Source Daunert et al., 2000
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
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Action-Potential Biosensor
Source Tonomura et al., 2006
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
33
Action-Potential Biosensor
(Side view)
Source Tonomura et al., 2006
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
34
Action-Potential Biosensor
Suction
Source Tonomura et al., 2006
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
35
Action-Potential Biosensor
Suction
Source Tonomura et al., 2006
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
36
Action-Potential Biosensor
Source Tonomura et al., 2006
Carbon N-T
E-DNA
BioFET
Whole Cell
Summary
Introduction
37
Summary

• Use of biomolecules in sensors offers
• Extreme sensitivity
• Flexibility of use
• Wide array of detection
• Universal application

Carbon N-T
E-DNA
BioFET
Introduction
Whole Cell
Summary
38
Summary

• But still maintains challenges of
• pH/Temperature sensitivity
• Degradation
• Repeatable use
• Regardless of challenges
• Biosensors will permeate future society

Carbon N-T
E-DNA
BioFET
Introduction
Whole Cell
Summary
39
References

• K McKimmie. Whats a Biosensor, Anyway?,
  Indiana Business Magazine, 2005, 49, 118-23.
• N Zimmerman. Chemical Sensors Market Still
  Dominating Sensors, Materials Management in
  Health Care, 2006, 2, 54.
• K Odenthal, J Gooding. An introduction to
  electrochemical DNA biosensors, Analyst, 2007,
  132, 603610.
• S V Lemeshko, T Powdrill, Y Belosludtsev, M
  Hogan, Oligonucleotides form a duplex with
  non-helical properties on a positively charged
  surface, Nucleic Acids Res., 2001, 29,
  30513058.
• F Ricci, R Lai, A Heeger, K Plaxco, J Sumner.
  Effect of Molecular Crowding on the Response of
  an Electrochemical DNA Sensor, Langmuir, 2007,
  23, 6827-6834.
• M Heller. DNA Microarray Technology, Annual
  Review of Biomedical Engineering, 2002, 4,
  129-153.
• E Boon, D Ceres, T Drummond, M Hill, J Barton,
  Mutation Detection by DNA electrocatalysis at
  DNA-modified electrodes, Nat. Biotechnol. 2000,
  18, 1096-1100.
• S Timur, U Anik, D Odaci, L Gorton, Development
  of a microbial biosensor based on carbon nanotube
  (CNT) modified electrodes, Electrochemistry
  Communications, 2007, 9, 1810-1815.
• K Besteman, J Lee, F Wiertz, H Heering, C Dekker.
  Enzyme-Coated Carbon Nanotubes as
• Single-Molecule Biosensors, Nano Letters, 2003,
  3, 6 727-730.
• R Chen, Y Zhang, D Wang, H Dai. Noncovalent
  Sidewall Functionalization of Single-Walled
  Carbon Nanotubes for Protein Immobilization, J.
  Am. Chem. Soc., 2001, 123, 16 3838 -3839.
• K Balasubramanian, M Burghard. Biosensors based
  on carbon nanotubes, Anal. Bioanal. Chem., 2005,
  385, 452-468.
• Hayes Horowitz, Student Manual for the Art of
  Electronics, Cambridge Univ. Press, 1989.
• I Hyungsoon, H Xing-Jiu, G Bonsang, C Yang-Kyu.
  A dielectric-modulated field-effect transistor
  for biosensing, Nature Nanotechnology, 2007, 2,
  430 434.
• D Therriault. Filling the Gap, Nature
  Nanotechnology, 2007, 2, 393 - 394.
• S Daunert, GBarrett, J Feliciano, R Shetty, S
  Shrestha, W Smith-Spencer. Genetically
  Engineered Whole-Cell Sensing Systems Coupling
  Biological Recognition with Reporter Genes,
  Chem. Rev. 2000, 100, 2705-2738.
• T Petänen, M Romantschuk. Measurement of
  bioavailability of mercury and arsenite using
  bacterial biosensors, Chemosphere, 2003, 50,
  409-413.
• F Roberto, J Barnes, D Bruhn. Evaluation of a
  GFP Reporter Gene Construct for Environmental
  Arsenic Detection., Talanta. 2002, 58,
  1181-188.
• W Tonomura, R Kitazawa, T Ueyama, H Okamura, S
  Konishi. Electrophysiological biosensor with
  Micro Channel Array for Sensing Signals from
  Single Cells, IEEE Sensors, 2006, 140-143.

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Questions?
Carbon N-T
E-DNA
BioFET
Introduction
Whole Cell
Summary