Bioanalytical chemistry. (2016)

• Record Type: Book
• Further Information: Note: Susan R. Mikkelsen, Eduardo Cortón.
• Authors: Mikkelsen, Susan R, 1960-; Cortón, Eduardo, 1962-
• Contents: 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 1.2.1. UV-Visible Absorbance 3 1.2.2. Turbidimetry (Light-Scattering) 5 1.2.3. Fluorescence 5 1.2.4. Chemiluminescence and Bioluminescence 7 1.3. Electrochemical Measurements 8 1.3.1. Potentiometry 10 1.3.2. Amperometry 10 1.3.3. Impedimetry 11 1.4. Radiochemical Measurements 12 1.4.1. Scintillation Counting 12 1.4.2. Geiger Counting 12 1.5. Surface Plasmon Resonance 13 1.6. Calorimetry 14 1.6.1. Differential Scanning Calorimetry (DSC) 15 1.6.2. Isothermal Titration Calorimetry (ITC) 16 1.7. Automation: Microplates, Multiwell Liquid Dispensers and Microplate Readers 16 1.8. Calibration of Instrumental Measurements 18 1.8.1. External Standards 18 1.8.2. Internal Standards 19 1.8.3. Standard Additions 20 1.9. Quantitative and Semi-Quantitative Measurements 21 1.9.1. Exact Concentration 21 1.9.2. Positive or Negative Result 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 2.2.1. Lowry Method 24 2.2.2. Smith (BCA) Method 24 2.2.3. Bradford Method 26 2.2.4. Ninhydrin-Based Assay 27 2.2.5. Other Protein Quantitation Methods 28 2.3. Total DNA 31 2.3.1. Diaminobenzoic Acid (DABA) Method 32 2.3.2. Diphenylamine (DPA) Method 32 2.3.3. Other Fluorimetric Methods 33 2.4. Total RNA 34 2.5. Total Carbohydrate 35 2.5.1. Ferricyanide Method 35 2.5.2. Phenol-Sulfuric Acid Method 36 2.5.3. 2-Aminothiophenol Method 36 2.5.4. Purpald Assay for Bacterial Polysaccharides 37 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 3.5.1. Simple One-Substrate Enzyme Kinetics 48 3.5.2. Experimental Determination of Michaelis-Menten Parameters 50 3.5.2.1. Eadie-Hofstee Method 50 3.5.2.2. Hanes Method 50 3.5.2.3. Lineweaver-Burk Method 51 3.5.2.4. Cornish-Bowden-Eisenthal Method 52 3.5.3. Comparison of Methods for the Determination of KM Values 52 3.5.4. One-Substrate, Two-Product Enzyme Kinetics 54 3.5.5. Two-Substrate Enzyme Kinetics 54 3.5.6. Examples of Enzyme-Catalyzed Reactions and their Treatment 56 3.5.7. Curve Fitting for Enzyme Kinetic Data 57 3.6. Enzyme Activators 58 3.7. Enzyme Inhibitors 59 3.7.1. Competitive Inhibition 60 3.7.2. Noncompetitive Inhibition 60 3.7.3. Uncompetitive Inhibition 62 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 4.5.1. Optical Detection 73 4.5.1.1. Absorbance 73 4.5.1.2. Fluorescence 75 4.5.1.3. Luminescence 77 4.5.1.4. Nephelometry 79 4.5.2. Electrochemical Detection 79 4.5.2.1. Amperometry 79 4.5.2.2. Potentiometry 80 4.5.2.3. Conductimetry 80 4.5.3. Other Detection Methods 81 4.5.3.1. Radiochemical 81 4.5.3.2. Manometry 81 4.5.3.3. Calorimetry 82 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 5.2.1. Nonpolymerizing Covalent Immobilization 91 5.2.1.1. Controlled-Pore Glass 92 5.2.1.2. Polysaccharides 93 5.2.1.3. Polyacrylamide 95 5.2.1.4. Acidic Supports 95 5.2.1.5. Anhydride Groups 96 5.2.1.6. Thiol Groups 97 5.2.2. Crosslinking with Bifunctional Reagents 97 5.2.3. Adsorption 98 5.2.4. Entrapment 99 5.2.5. Microencapsulation 100 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 6.5.1. Agglutination Reactions 124 6.5.2. Precipitation Reactions 126 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 7.3.1. Labeled-Antibody Methods 136 7.3.2. Labeled-Ligand Assays 136 7.3.3. Radioisotopes 139 7.3.4. Fluorophores 139 7.3.4.1. Indirect Fluorescence 140 7.3.4.2. Competitive Fluorescence 140 7.3.4.3. Sandwich Fluorescence 140 7.3.4.4. Fluorescence Excitation Transfer 140 7.3.4.5. Time-Resolved Fluorescence 141 7.3.5. Quantum Dots 142 7.3.6. Chemiluminescent Labels 143 7.3.7. Enzyme Labels 145 7.3.8. Lateral Flow Immunoassay 148 7.4. Homogeneous Immunoassays 149 7.4.1. Fluorescent Labels 149 7.4.1.1. Enhancement Fluorescence 149 7.4.1.2. Direct Quenching Fluorescence 150 7.4.1.3. Indirect Quenching Fluorescence 150 7.4.1.4. Fluorescence Polarization Immunoassay 151 7.4.1.5. Fluorescence Excitation Transfer 151 7.4.2. Enzyme Labels 152 7.4.2.1. Enzyme-Multiplied Immunoassay Technique 152 7.4.2.2. Substrate-Labelled Fluorescein Immunoassay 153 7.4.2.3. Apoenzyme Reactivation Immunoassay 153 7.4.2.4. Cloned Enzyme Donor Immunoassay 154 7.4.2.5. Enzyme Inhibitory Homogeneous Immunoassay 154 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 8.3.1. Natural Recognition Agents 171 8.3.2. Artificial Recognition Agents 172 8.4. Response of Enzyme-Based Biosensors 175 8.5. Examples of Biosensor Configurations 178 8.5.1. Ferrocene-Mediated Amperometric Glucose Sensor 178 8.5.2. Potentiometric Biosensor for Phenyl Acetate 180 8.5.3. Evanescent-Wave Fluorescence Biosensor for Bungarotoxin 181 8.5.4. Optical Biosensor for Glucose Based on Fluorescence Resonance Energy Transfer 183 8.5.5. Piezoelectric Sensor for Nucleic Acid Detection 184 8.5.6. Enzyme Thermistors 186 8.5.7. Fluorescence Sensor for Nitroaromatic Explosives Based on a Molecularly Imprinted Pol
• Edition: Second edition
• Publisher Details: Hoboken, New Jersey : John Wiley & Sons, Inc
• Publication Date: 2016
• Extent: 1 online resource
• Subjects: 572.36; Analytical biochemistry
• Languages: English
• ISBNs: 9781119057635
• Related ISBNs: 9781119057703
• Notes: Note: Description based on CIP data; resource not viewed.
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