Engineering the bioelectronic interface : applications to analyte biosensing and protein detection

The interfacing of man-made electronics with redox proteins and enzymes not only tells us a great deal about the levels of sophistication active in biology, but also paves the way to using it in derived sensory devices. Some of these have already had a profound impact on both clinical diagnostics and the quality of life enjoyed by those unfortunate enough to live with disease. Though much remains to be learnt about controlling and optimising these interfacial interactions, their potential uses are, if anything, growing. Written by leaders in the field, this is the only book to focus on the generation of biosensing interfaces with analyses and control at the molecular level. Some of these are enzyme based, others associated with the generation of surfaces for protein-protein recognition. Summaries of state-of-the-art investigations into the interfacing of structurally complex molecular species with electrode surfaces are included along with their design, analysis and potential application. Studies into the "wiring" of biomolecules to man-made surfaces through the use of delocalised "molecular wires" or carbon nanotubes are detailed as are the application of surface chemical and genetic engineering methods to the construction of robust, orientated biomolecular monolayers.

Author Notes

Dr Jason Davis (University of Oxford) has pioneered the application of scanning probe and fluorescence imaging technology to the analysis of bioelectrochemical interfaces; an understanding and control of which is clearly highly beneficial to the development of improved biosensing devices. From the early days of such studies, carried out with Professor Allen Hill FRS, his work has been refined to a level where single, active enzymes and proteins on electrode surfaces can be scrutinised under physiological and electrochemically-controlled conditions. Ground-breaking genetic methodologies are being applied to the generation of enzyme, protein or aptamer molecules which can be self-assembled, in an active form, on metallic electrode surfaces. His research group are also actively engaged in the assembly and construction of host-guest coordination complexes on surfaces, electroanalysis, molecular manipulation and molecular electronics. The group have published more than 80 papers in international journals.

Paul V. BernhardtJason J. Davis and Ben Peters and Yuki Hanyu and Wang XiItamar Willner and Ran Tel-Vered and Bilha WillnerJames F. Rusling and Xin Yu and Bernard S. Munge and Sang N. Kim and Fotios PapadimitrakopoulosH.A. Heering and G.W. CantersVikash R. Dodhia and Gianfranco GilardiJan Tkac and Jason J. DavisPaul Ko Ferrigno

Chapter 1 Communication with the Mononuclear Molybdoenzymes: Emerging Opportunities and Applications in Redox Enzyme Biosensors	p. 1
1.1 Introduction - the Three Mo Enzyme Families	p. 1
1.2 Mechanism	p. 2
1.3 Amperometric Biosensors	p. 3
1.4 Emerging Applications of Mo Enzymes in Sensing	p. 5
1.4.1 Xanthine Oxidase Family	p. 5
1.5 Sulfite Oxidase Family	p. 9
1.5.1 Sulfite Oxidoreductase	p. 10
1.6 DMSO Reductase Family	p. 15
1.6.1 DMSO Reductase	p. 15
1.6.2 Nitrate Reductase	p. 17
1.6.3 Arsenite Oxidase	p. 19
1.6.4 Chlorate and Perchlorate Reductase	p. 20
1.7 Conclusions	p. 20
References	p. 21
Chapter 2 Scanning Probe Analyses at the Bioelectronic Interface	p. 25
2.1 Introduction	p. 25
2.1.1 Scanning Probe Microscopy	p. 26
2.1.2 SPM Applications at the Biomolecular Interface	p. 35
2.1.3 Summary	p. 38
2.2 Bioelectronic Analyses	p. 39
2.2.1 Electrode Surface Considerations	p. 39
2.2.2 AFM Imaging Case Studies	p. 39
2.2.3 The Direct Imaging of Electrochemistry and Enzyme Activity	p. 41
2.2.4 Spectroscopic Assessment Electrodebiomolecule Electronic Coupling	p. 46
2.3 Summary	p. 49
References	p. 50
Chapter 3 Electrical Interfacing of Redox Enzymes with Electrodes by Surface Reconstitution of Bioelectrocatalytic Nanostructures	p. 56
3.1 Introduction	p. 56
3.2 Reconstituted Enzyme Electrodes in Monolayer Configurations	p. 59
3.3 Electrical Wiring of Redox Proteins with Electrodes by their Reconstitution on Cofactor-Functionalised Metallic Nanoparticles (NPs) or Carbon Nanotubes (CNTs)	p. 63
3.4 Reconstitution of apo-Enzymes in Thin Films of Redox Polymers	p. 70
3.5 Design of Electrically Contacted Enzyme Electrodes by the Crossing of Surface-confined Cofactor-enzyme Affinity Complexes	p. 72
3.6 Reconstituted Enzyme Electrodes for Biofuel Cell Applications	p. 82
3.7 Conclusions and Perspectives	p. 89
Acknowledgement	p. 90
References	p. 90
Chapter 4 Single-wall Carbon Nanotube Forests in Biosensors	p. 94
4.1 Unique Properties of Carbon Nanotubes	p. 94
4.1.1 Introduction	p. 94
4.1.2 Electrocatalytic Properties	p. 96
4.2 Biosensors Using Non-oriented Carbon Nanotube Electrodes	p. 96
4.3 Biosensors Utilising Vertically Aligned Carbon Nanotube Forests	p. 99
4.3.1 CNT Forest Fabrication	p. 99
4.3.2 Biosensor Applications of SWNT Forests	p. 107
4.4 Outlook for the Future	p. 112
References	p. 113
Chapter 5 Activating Redox Enzymes through Immobilisation and Wiring	p. 119
5.1 Introduction	p. 119
5.2 Protein Complexes	p. 120
5.2.1 Co-crystallisation	p. 120
5.2.2 Covalent Complexes	p. 122
5.3 Electron Transfer at Electrodes	p. 126
5.3.1 Voltammetry	p. 128
5.3.2 Chronoamperometry	p. 128
5.4 Surface Preparation	p. 132
5.4.1 Carbon	p. 332
5.4.2 Gold	p. 134
5.4.3 Other Methods	p. 134
5.5 Immobilisation	p. 136
5.5.1 Direct Immobilisation	p. 137
5.5.2 Wires	p. 139
5.5.3 Wiring Proteins	p. 143
5.6 Conclusion	p. 146
References	p. 146
Chapter 6 Cytochromes P450: Tailoring a Class of Enzymes for Biosensing	p. 153
6.1 Introduction	p. 153
6.2 Structure-function of Bacterial and Human Cytochromes P450	p. 155
6.3 The Need for Electrons: the Cytochrome P450 Catalytic Cycle	p. 158
6.4 Human Cytochromes P450 and Drug Metabolism	p. 161
6.5 Protein Engineering of P450s to Improve or Expand their Catalytic Properties	p. 165
6.5.1 Directed Evolution of Cytochrome P450 Enzymes	p. 166
6.5.2 Rational Design of Cytochrome P450 Enzymes	p. 167
6.6 Interfacing Cytochromes P450 to Electrodes	p. 171
6.6.1 Immobilisation on Unmodified Electrodes	p. 172
6.6.2 Immobilisation with Surfactants, Polymers and Gold Nanoparticles	p. 173
6.6.3 Immobilisation by Covalent Linkage on Gold Electrodes: Use of Spacers	p. 178
6.6.4 Protein Engineering to Control Protein Immobilisation and Catalytic Turnover on Electrode Surfaces	p. 180
6.7 Conclusions	p. 185
References	p. 186
Chapter 7 Label-free Field Effect Protein Sensing	p. 193
7.1 Interfacial Protein Detection	p. 193
7.2 Protein Microarrays	p. 194
7.2.1 Array Substrates	p. 194
7.2.2 Surface Chemistry and Immobilisation	p. 196
7.2.3 Capture Biomolecules	p. 198
7.2.4 Detection Tools	p. 200
7.2.5 Ultrasensitive Protein Detection	p. 204
7.3 Label-free Field Effect Protein Detection	p. 206
7.3.1 Field Effect Transistor (FET) based Protein Sensing	p. 207
7.3.2 Capacitance/Impedance Label-free Protein Sensing	p. 207
7.3.3 Nanoscale Devices for Label-free Field Effect Protein Sensing	p. 209
7.4 Conclusions	p. 213
References	p. 215
Chapter 8 Biological and Clinical Applications of Biosensors	p. 225
8.1 Biosensing for Pure Biological Research	p. 225
8.1.1 The Challenges of "Omics" and "Systems" Approaches	p. 225
8.1.2 Biological Complexity	p. 226
8.1.3 The Types of Device Required	p. 230
8.2 Biosensing for Clinical Applications	p. 231
8.2.1 The Clinical Problems-Diagnosis, Prognosis, Personalised Medicine	p. 231
8.2.2 Biosensors for Clinical Applications	p. 239
8.3 Further Reading	p. 240
References	p. 240
Subject Index	p. 243

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