Biosensors Christopher Byrd ENPM 808 B University of

Biosensors Christopher Byrd ENPM 808 B University of Maryland, College Park December 4, 2007

Outline n Introduction n 4 Specific Types of Biosensors ¡ Electrochemical (DNA) ¡ Carbon nanotube ¡ Bio. FET ¡ Whole Cell n Basic functionality n Benefits/Challenges n Summary n References

Introduction n Biosensor: Incorporation of a biomolecule in order to detect something Species to be detected Filter Recognition (analyte) Layer Introduction E-DNA Transducer Carbon N-T Electronics Bio. FET Whole Cell Signal Summary

Introduction n Biosensors ~ $3 B 90% → Glucose testing 8% - 10% increase in industry per year Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Electrochemical DNA Sensors n n Introduction Harnesses specificity of DNA Simple assembly Customizable Vast uses for small cost E-DNA Carbon N-T Bio. FET Whole Cell Summary

DNA Structure n n DNA structures---double helix 4 complementary bases: Adenine (A), Guanine (G), Thymine (T), and Cytosine (C) Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

DNA Specificity n Hydrogen bonding between base pairs n Stacking interaction between bases along axis of double-helix n Animation Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Principles of DNA biosensors n Nucleic acid hybridization (Target Sequence) (Hybridization) (Stable ds. DNA) ss. DNA (Probe) Source: http: //cswww. essex. ac. uk Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

E-DNA Sensor Structure “Stem-loop” s Gold electrode Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

E-DNA Sensor Structure Target “Stem-loop” s Gold electrode Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

E-DNA Sensor Structure (Open, extended) (Stem-loop) Source: Ricci et al. , Langmuir, 2007, 23, 6827 -6834 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Carbon Nanotube Biosensor Image: www. cnano-rhone-alpes. org Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Carbon Nanotube Biosensor n n One atom thick One nanometer diameter Ability to be functionalized Electrical conductivity as high as copper, thermal conductivity as high as diamond Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

CNT Biosensor Structure Succinimidyl ester Source: Chen et al. , 2001 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

CNT Uncoated vs. Coated Source: Chen et al. , 2001 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

CNT Biosensor Signal Detection O 2 Glucose Gluconic Acid H 2 O 2 e- Source: Besteman et al. , 2003 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

CNT Biosensor Signal Detection e- e- Effectively increases electrical current Source: Besteman et al. , 2003 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

CNT Biosensor Results 160 m. M 20 m. M Source: Besteman et al. , 2003 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Bio. FET n n Draws upon versatility of common electronic component (Field-Effect Transistor) Well understood expectations/results Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

FET + Drain Gate + + Insulator + + Source (Not conductive enough) (Electron Channel) - - Introduction E-DNA Carbon N-T - Bio. FET Whole Cell Summary

FET + Threshold Voltage Drain Gate + + Introduction E-DNA Carbon N-T Insulator + + Bio. FET - Source Whole Cell Summary

FET + Drain Gate ++ + ++ - - - E-DNA Source - - - Introduction - Insulator Carbon N-T - Bio. FET Whole Cell Summary

Bio. FET Source: Im et al. , 2007 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Bio. FET Results Gate (before) Source: Im et al. , 2007 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Bio. FET Results Gate (w/Gate complete (after. Biomolecule) etch, w/biotin) d Source: Im et al. , 2007 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Whole Cell Sensors Source: http: //www. whatsnextnetwork. com/technology/media/cell_adhesion. jpg Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Whole Cell Sensors n n n Harness normal genetic processes May detect dozens of pathogens Modifiable/customizable Reports bioavailability Temperature/p. H sensitive Short shelf-life Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Whole Cell Sensors Source: Daunert et al. , 2000 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Action-Potential Biosensor Source: Tonomura et al. , 2006 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Action-Potential Biosensor (Side view) Source: Tonomura et al. , 2006 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Action-Potential Biosensor Suction Source: Tonomura et al. , 2006 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Action-Potential Biosensor Source: Tonomura et al. , 2006 Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary

Summary n Use of biomolecules in sensors offers: ¡ ¡ Introduction Extreme sensitivity Flexibility of use Wide array of detection Universal application E-DNA Carbon N-T Bio. FET Whole Cell Summary

Summary n But still maintains challenges of: ¡ ¡ ¡ n p. H/Temperature sensitivity Degradation Repeatable use Regardless of challenges: ¡ Introduction Biosensors will permeate future society E-DNA Carbon N-T Bio. FET Whole Cell Summary

References n n n n n n K Mc. Kimmie. “What’s a Biosensor, Anyway? ”, Indiana Business Magazine, 2005, 49, 1: 18 -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, 603– 610. 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, 3051– 3058. 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, 1: 181 -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. R Leois, J Rae. “Low-noise patch-clamp techniques”, Meth. Enzym. 1998, 293: 218 -266. [1] A Vikas, C S Pundir. “Biosensors: Future Analytical Tools”, Sensors and Transducers, 2007, 2, 935 -944.

Questions? Introduction E-DNA Carbon N-T Bio. FET Whole Cell Summary