{"product_id":"bioanalytical-chemistry-isbn-9781118302545","title":"Bioanalytical Chemistry","description":"\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 \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  \u003cp\u003e\u003cstrong\u003eSusan R. Mikkelsen\u003c\/strong\u003e, PhD, is a Professor in the Department of Chemistry at the University of Waterloo, Ontario, Canada. She has presented at numerous conferences and is the author of over 35 peer-reviewed research articles and 115 presentations. She has organized and supervised a wide variety of bioanalytical research projects and has participated in local and international collaborations in this field. She is the co-author of the first edition of \u003cem\u003eBioanalytical Chemistry\u003c\/em\u003e. \u003c\/p\u003e\u003cp\u003e\u003cstrong\u003eEduardo Cortón\u003c\/strong\u003e, PhD, is Head of the Bioan?lisis and Biosensors Laboratory in the Biochemistry Department at the University of Buenos Aires, Argentina. He is also an Adjunct Professor in the Department of Biological Chemistry at the University of Buenos Aires as well as an active Independent Researcher at the National Council of Scientific and Technical Research (CONICET). He has published over 35 peer-reviewed research articles and presented at 80 conferences. He is the co-author of the first edition of \u003cem\u003eBioanalytical Chemistry\u003c\/em\u003e.  \u003c\/p\u003e\u003cp\u003eA timely, accessible survey of the multidisciplinary field of bioanalytical chemistry\u003c\/p\u003e \u003cp\u003eThe field of bioanalytical chemistry overlaps a diverse range of disciplines, including biotechnology, biopharmaceuticals, and diagnostics–all of which makes a comprehensive introduction to the subject even more essential for students and researchers. The Second Edition of \u003ci\u003eBioanalytical Chemistry\u003c\/i\u003e follows the same path of the first edition by providing a thorough introduction to bioanalytical chemistry for students and practitioners. In so doing, it brings together many of the techniques commonly used by biochemists and molecular biologists.\u003c\/p\u003e Along with many other topics, the text includes entire chapters on design and implementation of enzyme assays; mass spectrometry; and validation of new methods. Each chapter progresses from basic concepts to applications involving real samples, and ends with a set of problems, while an appendix contains selected answers. The significantly expanded new edition includes three new chapters and focuses on the more recent developments in bioanalytical chemistry while minimizing some of the techniques that are less relevant. The authors have limited mathematical derivations to those that are essential for a basic understanding of the principles underlying each method.\u003cbr\u003e\u003cbr\u003e \u003cp\u003eFeaturing many of the techniques used by biochemists and molecular biologists, \u003ci\u003eBioanalytical Chemistry\u003c\/i\u003e reviews all relevant aspects of using analytical methodology to solve biological problems. Each chapter in the book begins with coverage of basic concepts, then progresses to applications that involve real samples.\u003c\/p\u003e \u003cp\u003e    Bioanalytical Chemistry covers:\u003c\/p\u003e \u003cp\u003e    Biomimetic materials\u003c\/p\u003e \u003cp\u003e    Lab-on-chip devices\u003c\/p\u003e \u003cp\u003e    Spectroscopic methods for total protein, nucleic acids and carbohydrate\u003c\/p\u003e \u003cp\u003e    Structural and functional properties of antibodies\u003c\/p\u003e     Immunoassays for antigen or antibody quantitation\u003cbr\u003e\u003cbr\u003e \u003cp\u003e    Principles and proteomics applications of 1D and 2D electrophoresis\u003c\/p\u003e \u003cp\u003e    Centrifugation methods\u003c\/p\u003e \u003cp\u003e    Mass spectrometry of biomolecules\u003c\/p\u003e \u003cp\u003e    Quantitation of enzymes and their substrates\u003c\/p\u003e \u003cp\u003e    Design and implementation of high-throughput enzyme assays\u003c\/p\u003e \u003cp\u003e    Analytical methods\u003c\/p\u003e \u003cp\u003e    Validation of new methods\u003c\/p\u003e \u003cp\u003e    And much more\u003c\/p\u003e \u003cp\u003eThis indispensable guide features end-of-chapter problems and selected answers in the appendix, designed to help students assess their knowledge of each topic. Each chapter also includes suggestions for further reading.\u003c\/p\u003e \u003cp\u003eMore than any other reference, \u003ci\u003eBioanalytical Chemistry\u003c\/i\u003e captures the interdisciplinary scope of this emerging field–and uniquely prepares students for collaborative scientific work and research in industrial, government, and academic laboratories.\u003c\/p\u003e \u003cp\u003eSusan R. Mikkelsen, PhD, is a Professor in the Department of Chemistry at the University of Waterloo, Ontario, Canada. She has presented at numerous conferences and is the author of over 35 peer-reviewed research articles and 115 presentations. She has organized and supervised a wide variety of bioanalytical research projects and has participated in local and international collaborations in this field. She is the co-author of the first edition of \u003ci\u003eBioanalytical Chemistry\u003c\/i\u003e.\u003c\/p\u003e Eduardo Cortón, PhD, is Head of the Bioanálisis and Biosensors Laboratory in the Biochemistry Department at the University of Buenos Aires, Argentina.  He is also an Adjunct Professor in the Department of Biological Chemistry at the University of Buenos Aires as well as an active Adjunct Researcher at the National Council of Scientific and Technical Research (CONICET). He has published over 35 peer-reviewed research articles and presented at 80 conferences. He is the co-author of the first edition of \u003ci\u003eBioanalytical Chemistry\u003c\/i\u003e.","brand":"Wiley","offers":[{"title":"Default Title","offer_id":47988813201637,"sku":"NP9781118302545","price":106.0,"currency_code":"USD","in_stock":false}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1842\/7735\/files\/9781118302545.jpg?v=1761781691","url":"https:\/\/k12savings.com\/es\/products\/bioanalytical-chemistry-isbn-9781118302545","provider":"K12savings","version":"1.0","type":"link"}