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Bioanalytical chemistry / Susan R. Mikkelsen, Eduardo Corton. -- 2nd ed. -- Hoboken, New Jersey : John Wiley & Sons, Inc., c2016. – (58.1734/M636/2nd ed.) |
Contents
Preface to Second Edition
Preface to First Edition
Acknowledgments
1. Quantitative Instrumental Measurements
1.1. Introduction
1
1.2. Optical Measurements 2
1.3. Electrochemical Measurements 8
1.4. Radiochemical Measurements 12
1.5. Surface Plasmon Resonance 13
1.6. Calorimetry
14
1.7. Automation: Microplates, Multiwell Liquid
Dispensers and Microplate Readers 16
1.8. Calibration of Instrumental Measurements 18
1.9. Quantitative and Semi-Quantitative
Measurements 21
Suggested Reading
22
Problems
22
2. Spectroscopic Methods for the Quantitation of
Classes of Biomolecules 23
3.1. Introduction
41
3.2. Enzyme Nomenclature 42
3.3. Enzyme Commission Numbers 43
3.4. Enzymes in Bioanalytical Chemistry 45
3.5. Enzyme Kinetics 46
3.6. Enzyme Activators 58
3.7. Enzyme Inhibitors 59
3.8. Enzyme Units and Concentrations 62
Suggested Reading
64
References
64
Problems
64
4. Quantitation of Enzymes and Their Substrates
4.1. Introduction
67
4.2. Substrate Depletion or Product
Accumulation 68
4.3. Direct and Coupled Measurements 69
4.4. Classification of Methods 71
4.5. Instrumental Methods 73
4.6. High-Throughput Assays for Enzymes and
Inhibitors 82
4.7. Assays for Enzymatic Reporter Gene
Products 84
4.8. Practical Considerations for Enzymatic
Assays 85
Suggested Reading
86
References
86
Problems
87
5. Immobilized Enzymes
6. Antibodies
6.1. Introduction
117
6.2. Structural and Functional Properties of
Antibodies 118
6.3. Polyclonal and Monoclonal Antibodies 121
6.4. Antibody-Antigen Interactions 122
6.5. Analytical Applications of Secondary
Antibody-Antigen Interactions 124
Suggested Reading
129
References
129
Problems
129
7. Quantitative Immunoassays with Labels
7.1. Introduction
131
7.2. Labeling Reactions 132
7.3. Heterogeneous Immunoassays 134
7.4. Homogeneous Immunoassays 149
7.5. Evaluation of New Immunoassay Methods 155
Suggested Reading
160
References
160
Problems
161
8. Biosensors
8.1. Introduction
166
8.2. Biosensor Diversity and Classification 169
8.3. Recognition Agents 171
8.4. Response of Enzyme-Based Biosensors
8.5. Examples of Biosensor Configurations
8.6. Evaluation of Biosensor Perfomance 201
8.7. In Vivo Applications of Biosensors 202
Suggested Reading
207
References
207
Problems
209
9. Directed Evolution for the Design of
Macromolecular Reagents 210
9.1. Introduction
210
9.2. Rational Design and Directed Evolution 211
9.3. Generation of Genetic Diversity 214
9.4. Linking Genotype and Phenotype 217
9.5. Identification and Selection of Successful
Variants 221
9.6. Examples of Directed Evolution
Experiments 224
Suggested Reading
226
References
226
Problems
227
10. Image-Based Bioanalysis
10.1. Introduction
229
10.2. Magnification and Resolution 230
10.3. Optical Microscopy 231
10.4. Electron Microscopy 234
10.5. Scanning Tunneling Microscopy 237
10.6. Atomic Force Microscopy (AFM) 237
10.7. Scanning Electrochemical Microscopy
(SECM) 240
Suggested Reading
242
References
242
Problems
243
11. Principles of Electrophoresis
11.1. Introduction 244
11.2. Electrophoretic Support Media 248
11.3. Effect of Experimental Conditions
Onelectrophoretic Separations 254
11.4. Electric Field Strength Gradients 255
11.5. Pulsed Field Gel Electrophoresis (PFGE) 256
11.6. Detection of Proteins and Nucleic Acids After
Electrophoretic Separation 258
Suggested Reading
265
References
266
Problems
266
12. Applications of Zone Electrophoresis 268
12.1. Introduction 268
12.2. Determination of Protein Net Charge and
Molecular Weight Using PAGE 268
12.3. Determination of Protein Subunit Composition
and Subunit Molecular Weights
12.4. Molecular Weight of DNA by Agarose Gel
Electrophoresis 272
12.5. Identification of Isoenzymes 273
12.6. Diagnosis of Genetic (Inherited)
Disorders 274
12.7. DNA Fingerprinting and Restriction Fragment
Length Polymorphism 275
12.8. DNA Sequencing with the Maxam-Gilbert
Method 279
12.9. Immunoelectrophoresis 282
Suggested Reading
287
References
287
Problems
288
13. Isoelectric Focusing and 2D
Electrophoresis
290
14. Capillary Electrophoresis
15.1. Introduction 325
15.2. Sedimentation and Relative Centrifugal g
Force 325
15.3. Centrifugal Forces in Different Rotor
Types 327
15.4. Clearing Factor (K) 329
15.5. Density Gradients 330
15.6. Types of Centrifugation Techniques 333
15.7. Harvesting Samples 336
15.8. Analytical Ultracentrifugation 336
15.9. Selected Examples 342
Suggested Reading
346
References
346
Problems
347
16. Chromatography of Biomolecules
16.1. Introduction 349
16.2. Units and Definitions 350
16.3. Plate Theory of Chromatography 350
16.4. Rate Theory of Chromatography 351
16.5. Size Exclusion (Gel Filtration)
Chromatography 353
16.6. Stationary Phases For Size Exclusion
Chromatography 358
16.7. Affinity Chromatography 360
16.8. Ion-exchange Chromatography 368
Suggested Reading
374
References
374
Problems
375
17. Mass Spectrometry of Biomolecules
17.1. Introduction 377
17.2. Basic Description of the Instrumentation 379
17.3. Interpretation of Mass Spectra 386
17.4. Biomolecule Molecular Weight
Determination 388
17.5. Protein Identification 392
17.6. Protein-Peptide Sequencing
17.7. Nucleic Acid Applications
17.8. Bacterial Mass Spectrometry
17.9. Mass Spectrometry Imaging
Suggested Reading
401
References
401
Problems
402
18. Micro-TAS, Lab-on-a-Chip, and Microarray
Devices
18.1. Introduction 404
18.2. Device Fabrication Materials and Methods 405
18.3. Microfluidics 405
18.4. Detectors
407
18.5. Examples of Bioanalytical Devices 407
Suggested Reading
412
References
412
Problems
413
19. Validation of New Bioanalytical Methods
19.1. Introduction 414
19.2. Precision and Accuracy 415
19.3. Mean and Variance 416
19.4. Relative Standard Deviation and Other
Precision Estimators 417
19.5. Estimation of Accuracy 424
19.6. Qualitative (Screening) Assays 427
19.7. Examples of Validation Procedures 428
Suggested Reading
435
References
436
Answers to Selected Problems 437
Index 449