{"product_id":"bioanalytical-chemistry-9781118302545","title":"Bioanalytical Chemistry","description":"\u003cb\u003eBook Synopsis\u003c\/b\u003e\u003cbr\u003e\u003cp\u003eA timely, accessible survey of the multidisciplinary field of bioanalytical chemistry\u003c\/p\u003e \u003cul\u003e \u003cli\u003eProvides an all in one approach for both beginners and experts, from a broad range of backgrounds, covering introductions, theory, advanced concepts and diverse applications for each method\u003c\/li\u003e \u003c\/ul\u003e \u003cul\u003e \u003cli\u003eEach chapter progresses from basic concepts to applications involving real samples\u003c\/li\u003e \u003c\/ul\u003e \u003cul\u003e \u003cli\u003eIncludes three new chapters on Biomimetic Materials, Lab-on-Chip, and Analytical Methods\u003c\/li\u003e \u003cli\u003eContains end-of-chapter problems and an appendix with selected answers\u003c\/li\u003e \u003c\/ul\u003e\u003cbr\u003e\u003cbr\u003e\u003cb\u003eTable of Contents\u003c\/b\u003e\u003cbr\u003e\u003cp\u003ePreface to Second Edition xix\u003c\/p\u003e \u003cp\u003ePreface to First Edition xxi\u003c\/p\u003e \u003cp\u003eAcknowledgments xxiii\u003c\/p\u003e \u003cp\u003e\u003cb\u003e1. Quantitative Instrumental Measurements 1\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e1.1. Introduction 1\u003c\/p\u003e \u003cp\u003e1.2. Optical Measurements 2\u003c\/p\u003e \u003cp\u003e1.2.1. UV-Visible Absorbance 3\u003c\/p\u003e \u003cp\u003e1.2.2. Turbidimetry (Light-Scattering) 5\u003c\/p\u003e \u003cp\u003e1.2.3. Fluorescence 5\u003c\/p\u003e \u003cp\u003e1.2.4. Chemiluminescence and Bioluminescence 7\u003c\/p\u003e \u003cp\u003e1.3. Electrochemical Measurements 8\u003c\/p\u003e \u003cp\u003e1.3.1. Potentiometry 10\u003c\/p\u003e \u003cp\u003e1.3.2. Amperometry 10\u003c\/p\u003e \u003cp\u003e1.3.3. Impedimetry 11\u003c\/p\u003e \u003cp\u003e1.4. Radiochemical Measurements 12\u003c\/p\u003e \u003cp\u003e1.4.1. Scintillation Counting 12\u003c\/p\u003e \u003cp\u003e1.4.2. Geiger Counting 12\u003c\/p\u003e \u003cp\u003e1.5. Surface Plasmon Resonance 13\u003c\/p\u003e \u003cp\u003e1.6. Calorimetry 14\u003c\/p\u003e \u003cp\u003e1.6.1. Differential Scanning Calorimetry (DSC) 15\u003c\/p\u003e \u003cp\u003e1.6.2. Isothermal Titration Calorimetry (ITC) 16\u003c\/p\u003e \u003cp\u003e1.7. Automation: Microplates, Multiwell Liquid Dispensers and Microplate Readers 16\u003c\/p\u003e \u003cp\u003e1.8. Calibration of Instrumental Measurements 18\u003c\/p\u003e \u003cp\u003e1.8.1. External Standards 18\u003c\/p\u003e \u003cp\u003e1.8.2. Internal Standards 19\u003c\/p\u003e \u003cp\u003e1.8.3. Standard Additions 20\u003c\/p\u003e \u003cp\u003e1.9. Quantitative and Semi-Quantitative Measurements 21\u003c\/p\u003e \u003cp\u003e1.9.1. Exact Concentration 21\u003c\/p\u003e \u003cp\u003e1.9.2. Positive or Negative Result 21\u003c\/p\u003e \u003cp\u003eSuggested Reading 22\u003c\/p\u003e \u003cp\u003eProblems 22\u003c\/p\u003e \u003cp\u003e\u003cb\u003e2. Spectroscopic Methods for the Quantitation of Classes of Biomolecules 23\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e2.1. Introduction 23\u003c\/p\u003e \u003cp\u003e2.2. Total Protein 24\u003c\/p\u003e \u003cp\u003e2.2.1. Lowry Method 24\u003c\/p\u003e \u003cp\u003e2.2.2. Smith (BCA) Method 24\u003c\/p\u003e \u003cp\u003e2.2.3. Bradford Method 26\u003c\/p\u003e \u003cp\u003e2.2.4. Ninhydrin-Based Assay 27\u003c\/p\u003e \u003cp\u003e2.2.5. Other Protein Quantitation Methods 28\u003c\/p\u003e \u003cp\u003e2.3. Total DNA 31\u003c\/p\u003e \u003cp\u003e2.3.1. Diaminobenzoic Acid (DABA) Method 32\u003c\/p\u003e \u003cp\u003e2.3.2. Diphenylamine (DPA) Method 32\u003c\/p\u003e \u003cp\u003e2.3.3. Other Fluorimetric Methods 33\u003c\/p\u003e \u003cp\u003e2.4. Total RNA 34\u003c\/p\u003e \u003cp\u003e2.5. Total Carbohydrate 35\u003c\/p\u003e \u003cp\u003e2.5.1. Ferricyanide Method 35\u003c\/p\u003e \u003cp\u003e2.5.2. Phenol-Sulfuric Acid Method 36\u003c\/p\u003e \u003cp\u003e2.5.3. 2-Aminothiophenol Method 36\u003c\/p\u003e \u003cp\u003e2.5.4. Purpald Assay for Bacterial Polysaccharides 37\u003c\/p\u003e \u003cp\u003e2.6. Free Fatty Acids 37\u003c\/p\u003e \u003cp\u003eReferences 38\u003c\/p\u003e \u003cp\u003eProblems 39\u003c\/p\u003e \u003cp\u003e\u003cb\u003e3. Enzymes 41\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e3.1. Introduction 41\u003c\/p\u003e \u003cp\u003e3.2. Enzyme Nomenclature 42\u003c\/p\u003e \u003cp\u003e3.3. Enzyme Commission Numbers 43\u003c\/p\u003e \u003cp\u003e3.4. Enzymes in Bioanalytical Chemistry 45\u003c\/p\u003e \u003cp\u003e3.5. Enzyme Kinetics 46\u003c\/p\u003e \u003cp\u003e3.5.1. Simple One-Substrate Enzyme Kinetics 48\u003c\/p\u003e \u003cp\u003e3.5.2. Experimental Determination of Michaelis-Menten Parameters 50\u003c\/p\u003e \u003cp\u003e3.5.2.1. Eadie-Hofstee Method 50\u003c\/p\u003e \u003cp\u003e3.5.2.2. Hanes Method 50\u003c\/p\u003e \u003cp\u003e3.5.2.3. Lineweaver-Burk Method 51\u003c\/p\u003e \u003cp\u003e3.5.2.4. Cornish-Bowden-Eisenthal Method 52\u003c\/p\u003e \u003cp\u003e3.5.3. Comparison of Methods for the Determination of KM Values 52\u003c\/p\u003e \u003cp\u003e3.5.4. One-Substrate, Two-Product Enzyme Kinetics 54\u003c\/p\u003e \u003cp\u003e3.5.5. Two-Substrate Enzyme Kinetics 54\u003c\/p\u003e \u003cp\u003e3.5.6. Examples of Enzyme-Catalyzed Reactions and their Treatment 56\u003c\/p\u003e \u003cp\u003e3.5.7. Curve Fitting for Enzyme Kinetic Data 57\u003c\/p\u003e \u003cp\u003e3.6. Enzyme Activators 58\u003c\/p\u003e \u003cp\u003e3.7. Enzyme Inhibitors 59\u003c\/p\u003e \u003cp\u003e3.7.1. Competitive Inhibition 60\u003c\/p\u003e \u003cp\u003e3.7.2. Noncompetitive Inhibition 60\u003c\/p\u003e \u003cp\u003e3.7.3. Uncompetitive Inhibition 62\u003c\/p\u003e \u003cp\u003e3.8. Enzyme Units and Concentrations 62\u003c\/p\u003e \u003cp\u003eSuggested Reading 64\u003c\/p\u003e \u003cp\u003eReferences 64\u003c\/p\u003e \u003cp\u003eProblems 64\u003c\/p\u003e \u003cp\u003e\u003cb\u003e4. Quantitation of Enzymes and Their Substrates 67\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e4.1. Introduction 67\u003c\/p\u003e \u003cp\u003e4.2. Substrate Depletion or Product Accumulation 68\u003c\/p\u003e \u003cp\u003e4.3. Direct and Coupled Measurements 69\u003c\/p\u003e \u003cp\u003e4.4. Classification of Methods 71\u003c\/p\u003e \u003cp\u003e4.5. Instrumental Methods 73\u003c\/p\u003e \u003cp\u003e4.5.1. Optical Detection 73\u003c\/p\u003e \u003cp\u003e4.5.1.1. Absorbance 73\u003c\/p\u003e \u003cp\u003e4.5.1.2. Fluorescence 75\u003c\/p\u003e \u003cp\u003e4.5.1.3. Luminescence 77\u003c\/p\u003e \u003cp\u003e4.5.1.4. Nephelometry 79\u003c\/p\u003e \u003cp\u003e4.5.2. Electrochemical Detection 79\u003c\/p\u003e \u003cp\u003e4.5.2.1. Amperometry 79\u003c\/p\u003e \u003cp\u003e4.5.2.2. Potentiometry 80\u003c\/p\u003e \u003cp\u003e4.5.2.3. Conductimetry 80\u003c\/p\u003e \u003cp\u003e4.5.3. Other Detection Methods 81\u003c\/p\u003e \u003cp\u003e4.5.3.1. Radiochemical 81\u003c\/p\u003e \u003cp\u003e4.5.3.2. Manometry 81\u003c\/p\u003e \u003cp\u003e4.5.3.3. Calorimetry 82\u003c\/p\u003e \u003cp\u003e4.6. High-Throughput Assays for Enzymes and Inhibitors 82\u003c\/p\u003e \u003cp\u003e4.7. Assays for Enzymatic Reporter Gene Products 84\u003c\/p\u003e \u003cp\u003e4.8. Practical Considerations for Enzymatic Assays 85\u003c\/p\u003e \u003cp\u003eSuggested Reading 86\u003c\/p\u003e \u003cp\u003eReferences 86\u003c\/p\u003e \u003cp\u003eProblems 87\u003c\/p\u003e \u003cp\u003e\u003cb\u003e5. Immobilized Enzymes 90\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e5.1. Introduction 90\u003c\/p\u003e \u003cp\u003e5.2. Immobilization Methods 90\u003c\/p\u003e \u003cp\u003e5.2.1. Nonpolymerizing Covalent Immobilization 91\u003c\/p\u003e \u003cp\u003e5.2.1.1. Controlled-Pore Glass 92\u003c\/p\u003e \u003cp\u003e5.2.1.2. Polysaccharides 93\u003c\/p\u003e \u003cp\u003e5.2.1.3. Polyacrylamide 95\u003c\/p\u003e \u003cp\u003e5.2.1.4. Acidic Supports 95\u003c\/p\u003e \u003cp\u003e5.2.1.5. Anhydride Groups 96\u003c\/p\u003e \u003cp\u003e5.2.1.6. Thiol Groups 97\u003c\/p\u003e \u003cp\u003e5.2.2. Crosslinking with Bifunctional Reagents 97\u003c\/p\u003e \u003cp\u003e5.2.3. Adsorption 98\u003c\/p\u003e \u003cp\u003e5.2.4. Entrapment 99\u003c\/p\u003e \u003cp\u003e5.2.5. Microencapsulation 100\u003c\/p\u003e \u003cp\u003e5.3. Properties of Immobilized Enzymes 101\u003c\/p\u003e \u003cp\u003e5.4. Immobilized Enzyme Reactors 107\u003c\/p\u003e \u003cp\u003e5.5. Theoretical Treatment of Packed-Bed Enzyme Reactors 109\u003c\/p\u003e \u003cp\u003eSuggested Reading 113\u003c\/p\u003e \u003cp\u003eReferences 113\u003c\/p\u003e \u003cp\u003eProblems 114\u003c\/p\u003e \u003cp\u003e\u003cb\u003e6. Antibodies 117\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e6.1. Introduction 117\u003c\/p\u003e \u003cp\u003e6.2. Structural and Functional Properties of Antibodies 118\u003c\/p\u003e \u003cp\u003e6.3. Polyclonal and Monoclonal Antibodies 121\u003c\/p\u003e \u003cp\u003e6.4. Antibody-Antigen Interactions 122\u003c\/p\u003e \u003cp\u003e6.5. Analytical Applications of Secondary Antibody-Antigen Interactions 124\u003c\/p\u003e \u003cp\u003e6.5.1. Agglutination Reactions 124\u003c\/p\u003e \u003cp\u003e6.5.2. Precipitation Reactions 126\u003c\/p\u003e \u003cp\u003eSuggested Reading 129\u003c\/p\u003e \u003cp\u003eReferences 129\u003c\/p\u003e \u003cp\u003eProblems 129\u003c\/p\u003e \u003cp\u003e\u003cb\u003e7. Quantitative Immunoassays with Labels 131\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e7.1. Introduction 131\u003c\/p\u003e \u003cp\u003e7.2. Labeling Reactions 132\u003c\/p\u003e \u003cp\u003e7.3. Heterogeneous Immunoassays 134\u003c\/p\u003e \u003cp\u003e7.3.1. Labeled-Antibody Methods 136\u003c\/p\u003e \u003cp\u003e7.3.2. Labeled-Ligand Assays 136\u003c\/p\u003e \u003cp\u003e7.3.3. Radioisotopes 139\u003c\/p\u003e \u003cp\u003e7.3.4. Fluorophores 139\u003c\/p\u003e \u003cp\u003e7.3.4.1. Indirect Fluorescence 140\u003c\/p\u003e \u003cp\u003e7.3.4.2. Competitive Fluorescence 140\u003c\/p\u003e \u003cp\u003e7.3.4.3. Sandwich Fluorescence 140\u003c\/p\u003e \u003cp\u003e7.3.4.4. Fluorescence Excitation Transfer 140\u003c\/p\u003e \u003cp\u003e7.3.4.5. Time-Resolved Fluorescence 141\u003c\/p\u003e \u003cp\u003e7.3.5. Quantum Dots 142\u003c\/p\u003e \u003cp\u003e7.3.6. Chemiluminescent Labels 143\u003c\/p\u003e \u003cp\u003e7.3.7. Enzyme Labels 145\u003c\/p\u003e \u003cp\u003e7.3.8. Lateral Flow Immunoassay 148\u003c\/p\u003e \u003cp\u003e7.4. Homogeneous Immunoassays 149\u003c\/p\u003e \u003cp\u003e7.4.1. Fluorescent Labels 149\u003c\/p\u003e \u003cp\u003e7.4.1.1. Enhancement Fluorescence 149\u003c\/p\u003e \u003cp\u003e7.4.1.2. Direct Quenching Fluorescence 150\u003c\/p\u003e \u003cp\u003e7.4.1.3. Indirect Quenching Fluorescence 150\u003c\/p\u003e \u003cp\u003e7.4.1.4. Fluorescence Polarization Immunoassay 151\u003c\/p\u003e \u003cp\u003e7.4.1.5. Fluorescence Excitation Transfer 151\u003c\/p\u003e \u003cp\u003e7.4.2. Enzyme Labels 152\u003c\/p\u003e \u003cp\u003e7.4.2.1. Enzyme-Multiplied Immunoassay Technique 152\u003c\/p\u003e \u003cp\u003e7.4.2.2. Substrate-Labelled Fluorescein Immunoassay 153\u003c\/p\u003e \u003cp\u003e7.4.2.3. Apoenzyme Reactivation Immunoassay 153\u003c\/p\u003e \u003cp\u003e7.4.2.4. Cloned Enzyme Donor Immunoassay 154\u003c\/p\u003e \u003cp\u003e7.4.2.5. Enzyme Inhibitory Homogeneous Immunoassay 154\u003c\/p\u003e \u003cp\u003e7.5. Evaluation of New Immunoassay Methods 155\u003c\/p\u003e \u003cp\u003eSuggested Reading 160\u003c\/p\u003e \u003cp\u003eReferences 160\u003c\/p\u003e \u003cp\u003eProblems 161\u003c\/p\u003e \u003cp\u003e\u003cb\u003e8. Biosensors 166\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e8.1. Introduction 166\u003c\/p\u003e \u003cp\u003e8.2. Biosensor Diversity and Classification 169\u003c\/p\u003e \u003cp\u003e8.3. Recognition Agents 171\u003c\/p\u003e \u003cp\u003e8.3.1. Natural Recognition Agents 171\u003c\/p\u003e \u003cp\u003e8.3.2. Artificial Recognition Agents 172\u003c\/p\u003e \u003cp\u003e8.4. Response of Enzyme-Based Biosensors 175\u003c\/p\u003e \u003cp\u003e8.5. Examples of Biosensor Configurations 178\u003c\/p\u003e \u003cp\u003e8.5.1. Ferrocene-Mediated Amperometric Glucose Sensor 178\u003c\/p\u003e \u003cp\u003e8.5.2. Potentiometric Biosensor for Phenyl Acetate 180\u003c\/p\u003e \u003cp\u003e8.5.3. Evanescent-Wave Fluorescence Biosensor for Bungarotoxin 181\u003c\/p\u003e \u003cp\u003e8.5.4. Optical Biosensor for Glucose Based on Fluorescence Resonance Energy Transfer 183\u003c\/p\u003e \u003cp\u003e8.5.5. Piezoelectric Sensor for Nucleic Acid Detection 184\u003c\/p\u003e \u003cp\u003e8.5.6. Enzyme Thermistors 186\u003c\/p\u003e \u003cp\u003e8.5.7. Fluorescence Sensor for Nitroaromatic Explosives Based on a Molecularly Imprinted Polymer 187\u003c\/p\u003e \u003cp\u003e8.5.8. Immunosensor Microwell Arrays from Gold Compact Disks 188\u003c\/p\u003e \u003cp\u003e8.5.9. Nanoparticle-Enhanced Detection of Thrombin by SPR 190\u003c\/p\u003e \u003cp\u003e8.5.10. Environmental BOD and Toxicity Biosensors Based on Viable Cells 192\u003c\/p\u003e \u003cp\u003e8.5.11. Detection of Viruses using a Surface Acoustic Wave (SAW) Biosensor 193\u003c\/p\u003e \u003cp\u003e8.5.12. MEMS Microcantilever Biosensor for Virus Detection 196\u003c\/p\u003e \u003cp\u003e8.5.13. DNA Microarrays 198\u003c\/p\u003e \u003cp\u003e8.6. Evaluation of Biosensor Perfomance 201\u003c\/p\u003e \u003cp\u003e8.7. In Vivo Applications of Biosensors 202\u003c\/p\u003e \u003cp\u003e8.7.1. Biocompatible Materials 203\u003c\/p\u003e \u003cp\u003e8.7.2. Physiological Environment of the Human Body 203\u003c\/p\u003e \u003cp\u003e8.7.3. The Artificial Pancreas 205\u003c\/p\u003e \u003cp\u003e8.7.4. An Enzymatic Fuel Cell as a Component of an Implanted Biosensing System 205\u003c\/p\u003e \u003cp\u003e8.7.5. Other Examples of Implantable Biosensors 206\u003c\/p\u003e \u003cp\u003eSuggested Reading 207\u003c\/p\u003e \u003cp\u003eReferences 207\u003c\/p\u003e \u003cp\u003eProblems 209\u003c\/p\u003e \u003cp\u003e\u003cb\u003e9. Directed Evolution for the Design of Macromolecular Reagents 210\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e9.1. Introduction 210\u003c\/p\u003e \u003cp\u003e9.2. Rational Design and Directed Evolution 211\u003c\/p\u003e \u003cp\u003e9.3. Generation of Genetic Diversity 214\u003c\/p\u003e \u003cp\u003e9.3.1. Polymerase Chain Reaction and Error-Prone PCR 215\u003c\/p\u003e \u003cp\u003e9.3.2. DNA Shuffling 217\u003c\/p\u003e \u003cp\u003e9.4. Linking Genotype and Phenotype 217\u003c\/p\u003e \u003cp\u003e9.4.1. Cell Expression and Cell Surface Display (In vivo) 218\u003c\/p\u003e \u003cp\u003e9.4.2. Phage Display (In vivo) 218\u003c\/p\u003e \u003cp\u003e9.4.3. Ribosome Display (In vitro) 219\u003c\/p\u003e \u003cp\u003e9.4.4. mRNA-Peptide Fusion (In vitro) 220\u003c\/p\u003e \u003cp\u003e9.4.5. Microcompartmentalization (In vitro) 220\u003c\/p\u003e \u003cp\u003e9.5. Identification and Selection of Successful Variants 221\u003c\/p\u003e \u003cp\u003e9.5.1. Identification of Successful Variants Based on Binding Properties 222\u003c\/p\u003e \u003cp\u003e9.5.2. Identification of Successful Variants Based on Catalytic Activity 222\u003c\/p\u003e \u003cp\u003e9.6. Examples of Directed Evolution Experiments 224\u003c\/p\u003e \u003cp\u003e9.6.1. Directed Evolution of Galactose Oxidase 224\u003c\/p\u003e \u003cp\u003e9.6.2. α-Hemolysin Evolution 225\u003c\/p\u003e \u003cp\u003eSuggested Reading 226\u003c\/p\u003e \u003cp\u003eReferences 226\u003c\/p\u003e \u003cp\u003eProblems 227\u003c\/p\u003e \u003cp\u003e\u003cb\u003e10. Image-Based Bioanalysis 229\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e10.1. Introduction 229\u003c\/p\u003e \u003cp\u003e10.2. Magnification and Resolution 230\u003c\/p\u003e \u003cp\u003e10.3. Optical Microscopy 231\u003c\/p\u003e \u003cp\u003e10.3.1. The Compound Light Microscope 231\u003c\/p\u003e \u003cp\u003e10.3.2. The Confocal Microscope 231\u003c\/p\u003e \u003cp\u003e10.3.3. Sample Preparation 232\u003c\/p\u003e \u003cp\u003e10.3.4. General and Selective Stains 233\u003c\/p\u003e \u003cp\u003e10.3.5. Fluorescence In situ Hybridization 234\u003c\/p\u003e \u003cp\u003e10.3.6. Green Fluorescent Protein and its Analogues 234\u003c\/p\u003e \u003cp\u003e10.4. Electron Microscopy 234\u003c\/p\u003e \u003cp\u003e10.4.1. Principles and Instrumentation 234\u003c\/p\u003e \u003cp\u003e10.4.2. Sample Preparation 235\u003c\/p\u003e \u003cp\u003e10.4.3. Transmission Electron Microscopy (TEM) 236\u003c\/p\u003e \u003cp\u003e10.4.4. Scanning Electron Microscopy (SEM) 236\u003c\/p\u003e \u003cp\u003e10.5. Scanning Tunneling Microscopy 237\u003c\/p\u003e \u003cp\u003e10.5.1. Principles and Instrumentation 237\u003c\/p\u003e \u003cp\u003e10.5.2. Biological Applications 237\u003c\/p\u003e \u003cp\u003e10.6. Atomic Force Microscopy (AFM) 237\u003c\/p\u003e \u003cp\u003e10.6.1. Cantilevers and Operational Modes 237\u003c\/p\u003e \u003cp\u003e10.6.2. Samples and Substrates 239\u003c\/p\u003e \u003cp\u003e10.6.3. Biological Applications 239\u003c\/p\u003e \u003cp\u003e10.6.4. Four-Dimensional (4D) Scanning 240\u003c\/p\u003e \u003cp\u003e10.7. Scanning Electrochemical Microscopy (SECM) 240\u003c\/p\u003e \u003cp\u003e10.7.1. Principles and Instrumentation 240\u003c\/p\u003e \u003cp\u003e10.7.2. Samples and Substrates 241\u003c\/p\u003e \u003cp\u003e10.7.3. Biological Applications 241\u003c\/p\u003e \u003cp\u003eSuggested Reading 242\u003c\/p\u003e \u003cp\u003eReferences 242\u003c\/p\u003e \u003cp\u003eProblems 243\u003c\/p\u003e \u003cp\u003e\u003cb\u003e11. Principles of Electrophoresis 244\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e11.1. Introduction 244\u003c\/p\u003e \u003cp\u003e11.2. Electrophoretic Support Media 248\u003c\/p\u003e \u003cp\u003e11.2.1. Paper 248\u003c\/p\u003e \u003cp\u003e11.2.2. Starch Gels 249\u003c\/p\u003e \u003cp\u003e11.2.3. Polyacrylamide Gels 250\u003c\/p\u003e \u003cp\u003e11.2.4. Agarose Gels 254\u003c\/p\u003e \u003cp\u003e11.2.5. Polyacrylamide-Agarose Gels 254\u003c\/p\u003e \u003cp\u003e11.3. Effect of Experimental Conditions Onelectrophoretic Separations 254\u003c\/p\u003e \u003cp\u003e11.4. Electric Field Strength Gradients 255\u003c\/p\u003e \u003cp\u003e11.5. Pulsed Field Gel Electrophoresis (PFGE) 256\u003c\/p\u003e \u003cp\u003e11.6. Detection of Proteins and Nucleic Acids After Electrophoretic Separation 258\u003c\/p\u003e \u003cp\u003e11.6.1. Stains and Dyes 258\u003c\/p\u003e \u003cp\u003e11.6.2. Detection of Enzymes by Substrate Staining 260\u003c\/p\u003e \u003cp\u003e11.6.3. The Southern Blot 260\u003c\/p\u003e \u003cp\u003e11.6.4. The Northern Blot 262\u003c\/p\u003e \u003cp\u003e11.6.5. The Western Blot 262\u003c\/p\u003e \u003cp\u003e11.6.6. Detection of DNA Fragments on Membranes with DNA Probes 263\u003c\/p\u003e \u003cp\u003eSuggested Reading 265\u003c\/p\u003e \u003cp\u003eReferences 266\u003c\/p\u003e \u003cp\u003eProblems 266\u003c\/p\u003e \u003cp\u003e\u003cb\u003e12. Applications of Zone Electrophoresis 268\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e12.1. Introduction 268\u003c\/p\u003e \u003cp\u003e12.2. Determination of Protein Net Charge and Molecular Weight Using PAGE 268\u003c\/p\u003e \u003cp\u003e12.3. Determination of Protein Subunit Composition and Subunit Molecular Weights 270\u003c\/p\u003e \u003cp\u003e12.4. Molecular Weight of DNA by Agarose Gel Electrophoresis 272\u003c\/p\u003e \u003cp\u003e12.5. Identification of Isoenzymes 273\u003c\/p\u003e \u003cp\u003e12.6. Diagnosis of Genetic (Inherited) Disorders 274\u003c\/p\u003e \u003cp\u003e12.7. DNA Fingerprinting and Restriction Fragment Length Polymorphism 275\u003c\/p\u003e \u003cp\u003e12.8. DNA Sequencing with the Maxam-Gilbert Method 279\u003c\/p\u003e \u003cp\u003e12.9. Immunoelectrophoresis 282\u003c\/p\u003e \u003cp\u003eSuggested Reading 287\u003c\/p\u003e \u003cp\u003eReferences 287\u003c\/p\u003e \u003cp\u003eProblems 288\u003c\/p\u003e \u003cp\u003e\u003cb\u003e13. Isoelectric Focusing and 2D Electrophoresis 290\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e13.1. Introduction 290\u003c\/p\u003e \u003cp\u003e13.2. Carrier Ampholytes 291\u003c\/p\u003e \u003cp\u003e13.3. Modern IEF with Carrier Ampholytes 293\u003c\/p\u003e \u003cp\u003e13.4. Immobilized pH Gradients (IPGs) 296\u003c\/p\u003e \u003cp\u003e13.5. Two-Dimensional Electrophoresis 299\u003c\/p\u003e \u003cp\u003e13.6. Difference Gel Electrophoresis (DIGE) 301\u003c\/p\u003e \u003cp\u003eSuggested Reading 303\u003c\/p\u003e \u003cp\u003eReferences 303\u003c\/p\u003e \u003cp\u003eProblems 304\u003c\/p\u003e \u003cp\u003e\u003cb\u003e14. Capillary Electrophoresis 306\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e14.1. Introduction 306\u003c\/p\u003e \u003cp\u003e14.2. Electroosmosis 307\u003c\/p\u003e \u003cp\u003e14.3. Elution of Sample Components 308\u003c\/p\u003e \u003cp\u003e14.4. Sample Introduction 309\u003c\/p\u003e \u003cp\u003e14.5. Detectors for Capillary Electrophoresis 310\u003c\/p\u003e \u003cp\u003e14.5.1. Laser-Induced Fluorescence Detection 311\u003c\/p\u003e \u003cp\u003e14.5.2. Mass Spectrometric Detection 313\u003c\/p\u003e \u003cp\u003e14.5.3. Amperometric Detection 315\u003c\/p\u003e \u003cp\u003e14.5.4. Radiochemical Detection 318\u003c\/p\u003e \u003cp\u003e14.6. Capillary Polyacrylamide Gel Electrophoresis (C-PAGE) 319\u003c\/p\u003e \u003cp\u003e14.7. Capillary Isoelectric Focusing (CIEF) 321\u003c\/p\u003e \u003cp\u003eSuggested Reading 322\u003c\/p\u003e \u003cp\u003eReferences 323\u003c\/p\u003e \u003cp\u003eProblems 323\u003c\/p\u003e \u003cp\u003e\u003cb\u003e15. Centrifugation Methods 325\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e15.1. Introduction 325\u003c\/p\u003e \u003cp\u003e15.2. Sedimentation and Relative Centrifugal g Force 325\u003c\/p\u003e \u003cp\u003e15.3. Centrifugal Forces in Different Rotor Types 327\u003c\/p\u003e \u003cp\u003e15.3.1. Swinging-Bucket Rotors 327\u003c\/p\u003e \u003cp\u003e15.3.2. Fixed-Angle Rotors 328\u003c\/p\u003e \u003cp\u003e15.3.3. Vertical Rotors 328\u003c\/p\u003e \u003cp\u003e15.4. Clearing Factor (K) 329\u003c\/p\u003e \u003cp\u003e15.5. Density Gradients 330\u003c\/p\u003e \u003cp\u003e15.5.1. Materials Used to Generate a Gradient 331\u003c\/p\u003e \u003cp\u003e15.5.2. Constructing Pre-Formed and Self-Generated Gradients 331\u003c\/p\u003e \u003cp\u003e15.5.3. Redistribution of the Gradient in Fixed-Angle and Vertical Rotors 333\u003c\/p\u003e \u003cp\u003e15.6. Types of Centrifugation Techniques 333\u003c\/p\u003e \u003cp\u003e15.6.1. Differential Centrifugation 334\u003c\/p\u003e \u003cp\u003e15.6.2. Rate-Zonal Centrifugation 334\u003c\/p\u003e \u003cp\u003e15.6.3. Isopycnic Centrifugation 336\u003c\/p\u003e \u003cp\u003e15.7. Harvesting Samples 336\u003c\/p\u003e \u003cp\u003e15.8. Analytical Ultracentrifugation 336\u003c\/p\u003e \u003cp\u003e15.8.1. Instrumentation 337\u003c\/p\u003e \u003cp\u003e15.8.2. Sedimentation Velocity Analysis 338\u003c\/p\u003e \u003cp\u003e15.8.3. Sedimentation Equilibrium Analysis 341\u003c\/p\u003e \u003cp\u003e15.9. Selected Examples 342\u003c\/p\u003e \u003cp\u003e15.9.1. Analytical Ultracentrifugation for Quaternary Structure Elucidation 342\u003c\/p\u003e \u003cp\u003e15.9.2. Isolation of Retroviruses by Self-Generated Gradients 343\u003c\/p\u003e \u003cp\u003e15.9.3. Isolation of Lipoproteins from Human Plasma 344\u003c\/p\u003e \u003cp\u003e15.9.4. Centrifugal Microfluidic Analysis 344\u003c\/p\u003e \u003cp\u003eSuggested Reading 346\u003c\/p\u003e \u003cp\u003eReferences 346\u003c\/p\u003e \u003cp\u003eProblems 347\u003c\/p\u003e \u003cp\u003e\u003cb\u003e16.Chromatography of Biomolecules 349\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e16.1. Introduction 349\u003c\/p\u003e \u003cp\u003e16.2. Units and Definitions 350\u003c\/p\u003e \u003cp\u003e16.3. Plate Theory of Chromatography 350\u003c\/p\u003e \u003cp\u003e16.4. Rate Theory of Chromatography 351\u003c\/p\u003e \u003cp\u003e16.5. Size Exclusion (Gel Filtration) Chromatography 353\u003c\/p\u003e \u003cp\u003e16.6. Stationary Phases For Size Exclusion Chromatography 358\u003c\/p\u003e \u003cp\u003e16.6.1. Particulate Gels 358\u003c\/p\u003e \u003cp\u003e16.6.2. Monolithic Stationary Phases 360\u003c\/p\u003e \u003cp\u003e16.7. Affinity Chromatography 360\u003c\/p\u003e \u003cp\u003e16.7.1. Immobilization of Affinity Ligands 362\u003c\/p\u003e \u003cp\u003e16.7.2. Elution Methods 364\u003c\/p\u003e \u003cp\u003e16.7.3. Determination of Association Constants by High Performance Affinity Chromatography 364\u003c\/p\u003e \u003cp\u003e16.8. Ion-exchange Chromatography 368\u003c\/p\u003e \u003cp\u003e16.8.1. Retention Model for Ion-Exchange Chromatography of Polyelectrolytes 369\u003c\/p\u003e \u003cp\u003e16.8.2. Further Advances in Ion-Exchange Chromatography 374\u003c\/p\u003e \u003cp\u003eSuggested Reading 374\u003c\/p\u003e \u003cp\u003eReferences 374\u003c\/p\u003e \u003cp\u003eProblems 375\u003c\/p\u003e \u003cp\u003e\u003cb\u003e17. Mass Spectrometry of Biomolecules 377\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e17.1. Introduction 377\u003c\/p\u003e \u003cp\u003e17.2. Basic Description of the Instrumentation 379\u003c\/p\u003e \u003cp\u003e17.2.1. Soft Ionization Sources 379\u003c\/p\u003e \u003cp\u003e17.2.1.1. Fast Atom\/Ion Bombardment (FAB) 380\u003c\/p\u003e \u003cp\u003e17.2.1.2. Electrospray Ionization (ESI) 380\u003c\/p\u003e \u003cp\u003e17.2.1.3. Matrix-Assisted Laser Desorption\/Ionization (MALDI) 381\u003c\/p\u003e \u003cp\u003e17.2.2. Mass Analyzers 382\u003c\/p\u003e \u003cp\u003e17.2.3. Detectors 385\u003c\/p\u003e \u003cp\u003e17.3. Interpretation of Mass Spectra 386\u003c\/p\u003e \u003cp\u003e17.4. Biomolecule Molecular Weight Determination 388\u003c\/p\u003e \u003cp\u003e17.5. Protein Identification 392\u003c\/p\u003e \u003cp\u003e17.6. Protein-Peptide Sequencing 393\u003c\/p\u003e \u003cp\u003e17.7. Nucleic Acid Applications 397\u003c\/p\u003e \u003cp\u003e17.8. Bacterial Mass Spectrometry 398\u003c\/p\u003e \u003cp\u003e17.9. Mass Spectrometry Imaging 399\u003c\/p\u003e \u003cp\u003eSuggested Reading 401\u003c\/p\u003e \u003cp\u003eReferences 401\u003c\/p\u003e \u003cp\u003eProblems 402\u003c\/p\u003e \u003cp\u003e\u003cb\u003e18. Micro-TAS, Lab-on-a-Chip, and Microarray Devices 404\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e18.1. Introduction 404\u003c\/p\u003e \u003cp\u003e18.2. Device Fabrication Materials and Methods 405\u003c\/p\u003e \u003cp\u003e18.3. Microfluidics 405\u003c\/p\u003e \u003cp\u003e18.3.1. Fluid Transport 405\u003c\/p\u003e \u003cp\u003e18.3.2. Valves and Reservoirs 406\u003c\/p\u003e \u003cp\u003e18.3.3. Mixing and Sample Separation 406\u003c\/p\u003e \u003cp\u003e18.4. Detectors 407\u003c\/p\u003e \u003cp\u003e18.5. Examples of Bioanalytical Devices 407\u003c\/p\u003e \u003cp\u003e18.5.1. DNA Separation Using a Nanofence Array Microfluidic Device 408\u003c\/p\u003e \u003cp\u003e18.5.2. Two Dimensional Electrophoresis on a Microfluidic Chip 409\u003c\/p\u003e \u003cp\u003e18.5.3. Microfluidic Antibody Capture for Single-Cell Proteomics 410\u003c\/p\u003e \u003cp\u003e18.5.4. Multiplexed PCR Amplification and DNA Detection on a Microfluidic Chip 410\u003c\/p\u003e \u003cp\u003e18.5.5. Silicone Protein Separation Chip Based on a Grafted Ion-Exchange Polymer 411\u003c\/p\u003e \u003cp\u003e18.5.6. Circular, Biofunctionalized PEG Microchannels for Cell Adhesion Studies 411\u003c\/p\u003e \u003cp\u003eSuggested Reading 412\u003c\/p\u003e \u003cp\u003eReferences 412\u003c\/p\u003e \u003cp\u003eProblems 413\u003c\/p\u003e \u003cp\u003e\u003cb\u003e19. Validation of New Bioanalytical Methods 414\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e19.1. Introduction 414\u003c\/p\u003e \u003cp\u003e19.2. Precision and Accuracy 415\u003c\/p\u003e \u003cp\u003e19.3. Mean and Variance 416\u003c\/p\u003e \u003cp\u003e19.4. Relative Standard Deviation and Other Precision Estimators 417\u003c\/p\u003e \u003cp\u003e19.4.1. Distribution of Errors and Confidence Limits 418\u003c\/p\u003e \u003cp\u003e19.4.2. Linear Regression and Calibration 419\u003c\/p\u003e \u003cp\u003e19.4.3. Precision Profiles 420\u003c\/p\u003e \u003cp\u003e19.4.4. Limit of Quantitiation and Detection 421\u003c\/p\u003e \u003cp\u003e19.4.5. Linearizing Sigmoidal Curves (Four-Parameter Log-Logit Model) 422\u003c\/p\u003e \u003cp\u003e19.4.6. Effective Dose Method 423\u003c\/p\u003e \u003cp\u003e19.5. Estimation of Accuracy 424\u003c\/p\u003e \u003cp\u003e19.5.1. Standardization 424\u003c\/p\u003e \u003cp\u003e19.5.2. Matrix Effects 425\u003c\/p\u003e \u003cp\u003e19.5.2.1. Recovery 425\u003c\/p\u003e \u003cp\u003e19.5.2.2. Parallelism 426\u003c\/p\u003e \u003cp\u003e19.5.3. Interferences 426\u003c\/p\u003e \u003cp\u003e19.6. Qualitative (Screening) Assays 427\u003c\/p\u003e \u003cp\u003e19.6.1. Figures of Merit for Qualitative (Screening) Assays 427\u003c\/p\u003e \u003cp\u003e19.7. Examples of Validation Procedures 428\u003c\/p\u003e \u003cp\u003e19.7.1. Validation of a Qualitative Antibiotic Susceptibility Test 428\u003c\/p\u003e \u003cp\u003e19.7.2. Measurement of Plasma Homocysteine by Fluorescence Polarization Immunoassay (FPIA) Methodology 429\u003c\/p\u003e \u003cp\u003e19.7.3. Determination of Enzymatic Activity of β-Galactosidase 433\u003c\/p\u003e \u003cp\u003e19.7.4. Establishment of a Cutoff Value for Semi-Quantitative Assays for Cannabinoids 434\u003c\/p\u003e \u003cp\u003eSuggested Reading 435\u003c\/p\u003e \u003cp\u003eReferences 436\u003c\/p\u003e \u003cp\u003eAnswers to Selected Problems 437\u003c\/p\u003e \u003cp\u003eIndex 449\u003c\/p\u003e","brand":"John Wiley \u0026 Sons Inc","offers":[{"title":"Default Title","offer_id":49406848794967,"sku":"9781118302545","price":95.36,"currency_code":"GBP","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0817\/1739\/5799\/files\/9781118302545.jpg?v=1730497332","url":"https:\/\/bookcurl.com\/products\/bioanalytical-chemistry-9781118302545","provider":"Book Curl","version":"1.0","type":"link"}