Biosensors A Practical Approach

by Cooper, Jon; Cass, Tony

• ISBN13: 9780199638468
• ISBN10: 0199638462
• Edition: 2nd
• Format: Hardcover
• Copyright: 2004-05-06
• Publisher: Oxford University Press

Summary

Over the past 20 years, the field of biosensor research has had a significant impact in both laboratory research and the commercial sector. Over that period, biosensors have revolutionized the care and management of diabetes and have had important impacts in several other areas of clinical diagnostics. Europe, North America and Asia-Pacific have all seen the rise of small and medium sized companies seeking technical and application niches in the manufacture or use of biosensors. The current activity in both gene and protein 'biochips' can be seen as the latest set of tools that allow users who are not analytical science practitioners to make technically complex and reliable biological weapons and the need for their rapid and reliable detection will need to be met by devices that have many characteristics in common with biosensors. This book provides a practical introduction to the many skills needed in the highly interdisciplinary field of biosensor technology. Edited by two internationally renowned experts in this field, it draws together contributions from active researchers in Europe, North America and Asia who describe how to implement techniques as diverse as protein engineering, optical and electrochemical instrumentation, single cell electrophysiology, screen printing and numerical modelling in the context of producing biosensors for both laboratory and commercial applications. As well as the many detailed protocols and experimental tips, the book also offers an overview of current research in this area as well as pointers to its further directions. The diversity of topics covered means that it will be suitable both for those already active in the area who wish to expand their repertoire of experimental tools and for those who are just starting out in biosensors research.

Table of Contents

Protocol list	xi
Abbreviations	xiii
Contributors	xv
1 Redox hydrogel-based electrochemical biosensors
Adam Heller
1 Electron conducting redox polymers in biosensors
1.1 Relationship to mediator-based sensors	1	(1)
1.2 The electrocatalytic activity of redox hydrogels and the "wiring" of enzymes	2	(1)
1.3 Dependence of the diffusivity of electrons on crosslinking and the advantage of composite electrodes	2	(1)
1.4 The redox polymers and their electrochemistry	3	(1)
1.5 Crosslinkers and crosslinking	4	(1)
2 Enzyme electrodes	4	(3)
2.1 Electron transfer between enzyme and polymer redox centers	4	(2)
2.2 Microscopic homogeneity and salt effects in the redox polymer-enzyme system	6	(1)
2.3 Optimal compositions	6	(1)
2.4 Special matrices	7	(1)
3 Specific sensor examples	7	(12)
3.1 Amperometric and potentiometric biosensors of substrates of "wired" redox enzymes	7	(1)
3.2 Sensor measuring the turnover rate of hydrolytic and other non-redox enzymes	8	(2)
3.3 In vivo glucose sensors	10	(1)
3.4 Affinity sensors	10	(9)
2 Hybridization at oligonucleotide sensitive electrodes	19	(22)
Daren J. Caruana
1 Introduction	19	(1)
2 Function of oligonucleotide sensitive electrodes	20	(1)
3 Hybridization efficiency and sensitivity	21	(1)
4 Probe oligonucleotide structure and dynamics	22	(8)
4.1 Surface concentration	22	(1)
4.2 Probe length and orientation	23	(2)
4.3 Attachment of probe	25	(5)
5 Hybridization conditions	30	(6)
5.1 Temperature	31	(1)
5.2 Ionic strength	32	(1)
5.3 Base mismatch	33	(1)
5.4 Mass transport	34	(1)
5.5 Nonspecific adsorption	34	(2)
5.6 Other factors	36	(1)
6 Hybridization kinetics	36	(2)
7. Summary	38	(3)
3 Screen-printing methods for biosensor production	41	(18)
Xian-En Zhang
1 Introduction	41	(1)
2 Screen-printing technology	42	(9)
2.1 Materials and methods	42	(5)
2.2 Apparatus	47	(1)
2.3 Printing patterns	48	(1)
2.4 Printing process	49	(2)
3 Applications	51	(4)
3.1 Clinical diagnosis	51	(1)
3.2 Food analysis bioprocess control	52	(1)
3.3 Environmental monitoring	52	(2)
3.4 Other approaches	54	(1)
4 Conclusion	55	(4)
4 Kinetic modeling for biosensors	59	(38)
Philip Bartlett and Chee-Seng Toh
1 Introduction	59	(10)
1.1 The purpose and practice of modeling	59	(1)
1.2 Enzyme kinetics	60	(3)
1.3 Basic electrochemistry	63	(6)
2 Modeling	69	(20)
2.1 The flux diagram for the membrane/enzyme/electrode	70	(1)
2.2 Simplifying assumptions	70	(1)
2.3 The flux equations	71	(2)
2.4 Solution of flux equations	73	(5)
2.5 Deriving a complete kinetic model	78	(3)
2.6 Experimental verification of approximate analytical kinetic models	81	(1)
2.7 Numerical simulation methods	82	(7)
3 Applications	89	(1)
4 Kinetic modeling in other types of biosensors	89	(3)
4.1 Potentiometric enzyme electrodes	90	(1)
4.2 Optical and photometric biosensors	90	(1)
4.3 Immunosensors	91	(1)
5 Conclusions	92	(5)
List of symbols	92	(5)
5 Bio-, chemi-, and electrochemiluminescence for fiber-optic biosensors	97	(12)
Loic J. Blum and Pierre R. Coulet
1 Introduction	97	(1)
2 Design of the biosensor	97	(8)
2.1 Optical waveguide	97	(1)
2.2 Setup	98	(2)
2.3 Light-emitting reactions	100	(1)
2.4 Preparation of the sensing layer	101	(4)
3 Examples of determinations with the luminescence sensors	105	(3)
3.1 ATP determination	105	(1)
3.2 NADH determination	105	(1)
3.3 Extension to other analytes using dehydrogenases as auxiliary enzymes	105	(1)
3.4 H2O2 determination	106	(1)
3.5 Extension to other analytes involving H2O2 detection	107	(1)
4 Concluding remarks	108	(1)
6 Determination of metal ions by fluorescence aniostropy: A practical biosensing approach	109	(22)
Richard Thompson, Badri Maliwal, Hui Hui Zeng, and Michele Loetz Cramer
1 Introduction and rationale	109	(2)
1.1 Why fluorescence anisotropy to determine metal ions?	109	(2)
2 Theory of anisotropy-based determination of metal ions	111	(3)
2.1 "Reagent" approaches	111	(1)
2.2 "Reagentless" approach	112	(2)
3 Fluorescent aryl sulfonamides for zinc(II) determination	114	(1)
4 Removal of zinc from carbonic anhydrase (CA)	115	(2)
5 Avoidance of metal ion contamination	117	(2)
6 Determination of Zn using a reagent approach	119	(4)
7 Determination of Cu and other ions by using a reagentless approach	123	(1)
8 Calibration of anisotropy	124	(7)
7 Fluorescence-based fiber-optic biosensors	131	(22)
David R. Walt, Caroline L. Schauer, Shannon E. Stitzel, Michael S. Fleming, and Jason R. Epstein
1 Introduction	131	(3)
1.1 Fiber polishing	133	(1)
2 Single-analyte detection using an enzymatic sensing layer	134	(2)
2.1 Enzymatic sensing layer	134	(1)
2.2 PAN gel immobilization	135	(1)
3 Multi-analyte arrays	136	(15)
3.1 Immobilization via polymer photodeposition	136	(2)
3.2 Microwell array platform preparation	138	(8)
3.3 Live-cell array fabrication	146	(5)
4 Conclusions	151	(2)
8 Functional analysis of ion channels: Planar patch clamp and impedance spectroscopy of tethered lipid membranes	153	(32)
Michael Mayer, Samuel Terrettaz, Laurent Giovangrandi, Thierry Stora, and Horst Vogel
1 Introduction	153	(1)
2 Planar patch clamp	154	(14)
2.1 Concept of patch clamp on a chip	154	(3)
2.2 Formation of planar bilayers on a chip	157	(8)
2.3 Chip-based planar bilayers: single-channel measurements of alamethicin pores	165	(3)
3 Impedance spectroscopy of tethered lipid membranes	168	(17)
3.1 Basics of impedance spectroscopy	168	(2)
3.2 Measuring technique and electrochemical cell	170	(1)
3.3 Hybrid lipid layer	170	(4)
3.4 Tethered lipid bilayers	174	(3)
3.5 Lipid bilayer tethered via surface-attached proteins	177	(2)
3.6 Highly insulating tethered lipid bilayers for single-channel experiments	179	(6)
9 Protein engineering for biosensors	185	(56)
Gianfranco Gilardi
1 Introduction	185	(2)
2 Rational protein engineering	187	(35)
2.1 Modeling and calculations on protein structures	188	(11)
2.2 Site-directed mutagenesis	199	(23)
3 Directed evolution	222	(12)
3.1 Random mutagenesis: error prone PCR	224	(2)
3.2 Recombination: DNA shuffling	226	(4)
3.3 Functional screening of the library	230	(4)
4 Functional characterization of the mutants	234	(1)
5 Other aspects of protein engineering	234	(4)
6 Concluding remarks	238	(3)
Index	241