ÊéÃû bioanalytical chemistry
×÷ÕßAndreas Manz
Nicole Pamme
Dimitri lossif idis
Ò³Êý 217
ÓïÑÔ Ó¢Óï
³ö°æÉç Imperial College Press
ISBN 1-86094-370-5
Ŀ¼Preface ix
List of Abbreviations xi
Chapter 1 Biomolecules 1
1.1 Amino Acids, Peptides and Proteins 1
1.1.1 Amino Acids 2
1.1.2 Peptides and Proteins 7
1.2 Nucleic Acids 14
1.2.1 The Structure of Nucleic Acids 15
1.2.2 Synthesis of Proteins 20
1.3 Biomolecules in Analytical Chemistry 22
1.3.1 Classical Analytical Chemistry 22
1.3.2 Limitations of Classical Analytical Chemistry 22
1.3.3 Bioanalytical Chemistry 23
Chapter 2 Chromatography 29
2.1 The Principle of Chromatography 29
2.2 Basic Chromatographic Theory 31
2.3 Application of Liquid Chromatography for Bioanalysis 34
2.3.1 Reversed Phase Liquid Chromatography (RP-LC) 34
2.3.2 Ion Exchange Chromatography (IEC) 37
2.3.3 Affinity Chromatography 40
2.3.4 Size Exclusion Chromatography (SEC) 42
Chapter 3 Electrophoresis 47
3.1 Principle and Theory of Electrophoresis 48
3.1.1 Electrophoretic Mobility 49
3.1.2 Joule Heating 50
3.1.3 Electroosmotic Flow (EOF) 50
3.1.4 Separation Efficiency and Resolution 54
3.2 Gel Electrophoresis (GE) 56
3.2.1 Instrumentation for Gel Electrophoresis 57
3.2.2 Modes of Gel Electrophoresis 63
3.2.3 Sodium Dodecyl Sulphate¨CPolyacrylamide Gel
Electrophoresis (SDS¨CPAGE) 63
v
vi Contents
3.2.4 Isoelectric Focussing (IEF) 64
3.2.5 Two-Dimensional Gel Electrophoresis (2D-GE) 67
3.3 Capillary Electrophoresis (CE) 69
3.3.1 Capillary Electrophoresis Instrumentation 70
3.3.2 Capillary Zone Electrophoresis (CZE) 75
3.3.3 Capillary Isoelectric Focussing (CIEF) 76
3.3.4 Micellar Electrokinetic Chromatography (MEKC) 77
3.3.5 Capillary Gel Electrophoresis (CGE) 82
Chapter 4 Mass Spectrometry 85
4.1 The Principle of Mass Spectrometry 85
4.1.1 Ionisation 86
4.1.2 Mass Analyser 86
4.1.3 Detector 87
4.2 Matrix Assisted Laser Desorption Ionisation ¨C Time of Flight
Mass Spectrometry (MALDI-TOF/MS) 87
4.2.1 Ionisation Principle 87
4.2.2 Mass Analysis in Time-of-Flight Analyser 90
4.2.3 Detection of Ions 92
4.2.4 Resolution 92
4.2.5 Sample Pretreatment 93
4.2.6 Applications of MALDI 94
4.3 Electrospray Ionisation Mass Spectrometry (ESI-MS) 97
4.3.1 Ionisation Principle 98
4.3.2 ESI ¨C Source and Interface 99
4.3.3 Quadrupole Analyser 100
4.3.4 Applications of ESI-MS 101
Chapter 5 Molecular Recognition:
Bioassays, Biosensors, DNA-Arrays and
Pyrosequencing 109
5.1 Bioassays 110
5.1.1 Antibodies 111
5.1.2 Antigens 113
5.1.3 Antibody-Antigen Complex Formation 114
5.1.4 Assay Formats 115
5.1.5 Home Pregnancy Test 120
5.1.6 Enzyme Immunoassays (EI and ELISA) 121
5.2 Biosensors 125
5.2.1 Bioreceptors 126
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