Laboratory of Biosensors and Bioelectronics

Lecture #12: Nano-Biosensors

Objectives
Learn about new biosensing concepts that are based on nanotechnology Understand the principles of nanoparticle and nanowire based biosensing

Content
Introduction to Nanobiotechnology Nano-Biosensing Ion-channel Based Sensing N Nano-optics for bi ti f biosensing i DNA as a tool in nanotechnology Conductivity of DNA Nanowire Biosensing Principles of localized surface plasmon resonance sensing (Takumi Sannomiya)

Building blocks of life

Organs Tissues Cells C ll
nanometer r

Organelles p Macromolecule Complexes Macromolecules: proteins, DNA, lipids

The Birthday of Nanotechnology

December 29, 1959. Theres plenty of room in th b tt ! i the bottom!

Richard P. Feynman

1000 Encyclopedia Britannica on an A4

Nanobiotechnology

What is Nanobiotechnology?
Nanobiotechnology is the application of nanotechnology to life sciences. It includes: th application of nanoscale tools to biological the li ti f l t l t bi l i l systems the use of biological templates to create p nanoscale products
Nature Biotechnology 21:1137-1143 (2003)

Why Nanobiotechnology? Mother Nature did it first: A cell is a warehouse of nanoscale machines that function in a highly controlled manner. Biology can teach the physical world of electronics, materials science computing and manufacturing science, manufacturing. There exists biological analogues of conventional functional devices. We have the tools now to do it.

Nanobiotechnology Applications
Nanomedicine Nanobiosensors (biochips) Nanofluidics N fl idi Molecular Self-Assembly Intelligent Drug Delivery Systems Nanomachines

Hype or Hope?

Nanobio Promises: Science? Fiction?

Biosensing Concept

Enzymatic Biosensors - Principles and Applications. D. Grieshaber, R. MacKenzie, J. Vrs, and E. Reimhult, Sensors, 8(3); 1400-1458, 2008.

How can nanotechnology help?

Lets see some real examples!

Nanobiosensors: Ion-Channel Based Sensing

B.A. Cornell et al. Nature 387, 580-583 (1997)

Concept of Ion-Channel Based Sensing

Gramicidin dimer can form

Dimer formation is not possible

Current is detected through the channel

Current stops

Ion-Channel Based Biosensing

Ferritin

IgG

Streptavidin

Thyroid Stimulating Hormone

Gramicidin

The Gramicidin Dimer in the Membrane

Analyte Detection Examples

thyroid - stimulating hormone

Ion-Channel Based Sensing: Competitive Binding Configuration

Dimer formation is not possible

Gramicidin dimer can form

No N current is detected ti d t t d

Current flow starts C t fl t t through the channel

Example for a Competitive Assay

Addition of biotin-antibody blocks current

Adding co pe o restores cu e dd g competitor es o es current

Building-up the Surface Architecture

Immobilized gramicidin monomer Half-membrane spanning tethered lipids Spacers Lipids Biotinylated gramicidin monomer Half-membrane spanning mobile lipids Biotinylated lipids Streptavidin linker Biotinylated receptors

B.A. Cornell et al. Nature 387, 580-583 (1997)

Enhancing Sensitivity

AMBRI Sensor chip

http://www.ambri.com.au

Functionalization of array elements by spotting

Spotting

Applications
Point of care Point-of-care diagnostics Drug discovery Bacteriological detection B t i l i l d t ti Veterinary diagnostics Food testing g Environmental monitoring

Nanobiosensors: Transmission Plasmon Biosensor

Au A

Ag A

100nm

100nm

Color of Nanoparticles
Gold Nanoparticles Quantum Dots

4nm

50nm

Old-New Discovery

Absorption of Immobilized Particles

Protein Sensing with TPB

Au Ag

Anal. Chem., 75 (24), 6894 -6900, 2003.

Surface Ag Nanostructures

ADDL=amyloid -derived diffusible ligands

Nano Lett. 2004, 4, (6), 1029-1034

Immunosensing Performance

Nano Lett. 2004, 4, (6), 1029-1034

Optical Properties of Nanoholes and Disks

Nano Letters, 4 (6), 1003 -1007, 2004.

Size Dependence of Absorption Spectra of Single Nanoholes

Nano Letters, 4 (6), 1003 -1007, 2004.

Refractive Index Sensitivity

Nano Letters, 4 (6), 1003 -1007, 2004.

Dependence on the Density of Holes

Nano Letters, 4 (6), 1003 -1007, 2004.

Advantages
Simple absorption measurement Compatibility with array format Label-free technique

Applications
High through-put screening Interaction analysis

Problems
Sensitivity Theory and interpretation

DNA in Nanotechnology

Folding DNA to create nanoscale shapes and patterns Paul W. K. Rothemund, Nature 440, 297-302 (16 March 2006)

Folding DNA to create nanoscale shapes and patterns Paul W. K. Rothemund, Nature 440, 297-302 (16 March 2006)

DNA Nanotemplating

DNA-Templated Self-Assembly of Metallic Nanocomponent Arrays on a Surface NANO LETTERS, 2004, 4(12): 2343-2347

Summary
Nanobiosensors: Ion-Channel Based Sensing

Nanobiosensors: N N b Nanowire Arrays

B.A. Cornell et al. Nature 387, 580-583 (1997)

Nat. Biotechnol. 23, 1294-1301 (2005)

Nanobiosensors: Transmission Plasmon Biosensor

Au Ag

Immobilization of Single Vesicles on Nanopatterns

100nm

100nm

Angewandte Chemie International Edition 42, (45), 5580-5583, 2003

Nanobio Promises: Science? Fiction?

Program of the Exercise Class

Conductivity of DNA Nanowire Sensing Principle Nanooptical (plasmonic) sensing (Takumi Sannomiya)

Conductivity of DNA

Insulating Semiconducting Conducting

R. G. Endres et al, REVIEWS OF MODERN PHYSICS, VOLUME 76, JANUARY 2004

Reasons for Uncertainty

Contact Sequence specificity B kb Backbone and DNA environment d i t

Contact Examples
Using top-down techniques Or Carbon Nanotubes

I-V Curves of DNA Sequences

matching sequences (1,4)

Guo X et al, Nature Nanotechnology 3, 163 - 167 (2008)

mismatching sequences (2,3,5,6)

Semiconducting M-DNA

Calculated Co-DNA performance

Nokhrin S et al, Nanotechnology 18 (2007) 095205

Laboratory of Biosensors and Bioelectronics

Nano-Optical Sensing p g (with Localized Surface Plasmons)

Takumi Sannomiya