Bioanalytical Chemistry

19.04.2016

Englisch

9781118302545

John Wiley & Sons

Nein

Susan R. Mikkelsen

488

2

Gebundene Ausgabe

Bioanalytical Chemistry, Bioanalytik, Bioanalytische Chemie, Biochemie, Biochemie u. Chemische Biologie, Biochemistry (Chemical Biology), Biowissenschaften, Chemie, Chemistry, Life Sciences, Molecular Microbiology, Molekulare Mikrobiologie

PNF - Analytische Chemie

Preface to Second Edition xix Preface to First Edition xxi Acknowledgments xxiii 1. Quantitative Instrumental Measurements 1 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 2.1. Introduction 23 2.2. Total Protein 24 2.3. Total DNA 31 2.4. Total RNA 34 2.5. Total Carbohydrate 35 2.6. Free Fatty Acids 37 References 38 Problems 39 3. Enzymes 41 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 67 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 90 5.1. Introduction 90 5.2. Immobilization Methods 90 5.3. Properties of Immobilized Enzymes 101 5.4. Immobilized Enzyme Reactors 107 5.5. Theoretical Treatment of Packed-Bed Enzyme Reactors 109 Suggested Reading 113 References 113 Problems 114 6. Antibodies 117 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 131 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 166 8.1. Introduction 166 8.2. Biosensor Diversity and Classification 169 8.3. Recognition Agents 171 8.4. Response of Enzyme-Based Biosensors 175 8.5. Examples of Biosensor Configurations 178 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 229 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 244 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 270 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 13.1. Introduction 290 13.2. Carrier Ampholytes 291 13.3. Modern IEF with Carrier Ampholytes 293 13.4. Immobilized pH Gradients (IPGs) 296 13.5. Two-Dimensional Electrophoresis 299 13.6. Difference Gel Electrophoresis (DIGE) 301 Suggested Reading 303 References 303 Problems 304 14. Capillary Electrophoresis 306 14.1. Introduction 306 14.2. Electroosmosis 307 14.3. Elution of Sample Components 308 14.4. Sample Introduction 309 14.5. Detectors for Capillary Electrophoresis 310 14.6. Capillary Polyacrylamide Gel Electrophoresis (C-PAGE) 319 14.7. Capillary Isoelectric Focusing (CIEF) 321 Suggested Reading 322 References 323 Problems 323 15. Centrifugation Methods 325 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 349 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 377 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 393 17.7. Nucleic Acid Applications 397 17.8. Bacterial Mass Spectrometry 398 17.9. Mass Spectrometry Imaging 399 Suggested Reading 401 References 401 Problems 402 18. Micro-TAS, Lab-on-a-Chip, and Microarray Devices 404 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 414 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

250 mm

15 cm