Kinetics Guide - Binding kinetics with the WAVEsystem

All WAVEsystem products have been designed with user experience in mind. At the same time, they are highly sophisticated bioanalytical instruments requiring skilled user input. 

To help you get the most from your product, we have combined the wealth of our internal knowledge and experience in one practical reference manual. Our experts guide you through the main principles underlying the technologies, techniques and applications. We provide information, advice and tips on experimental design and data analysis. A separate troubleshooting section covers the situations and questions encountered most often by our customer support teams.

Whether you read it from cover to cover, refer to it for specific questions, or keep it as a handy reminder of equations and protocols, we hope that this manual will support you in your work and contribute to results that accelerate discovery.

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What's in the guide?
Content Page
1. Welcome 5
2. Introduction 7
2.1 Introduction to label-free interaction analysis 8
     2.1.1 Definitions of label-free terminology 12
     2.1.2 Principles of label-free analysis 16
2.2 The Creoptix WAVEsystem 19
     2.2.1 GCI technology 19
     2.2.3 Non-clog microfluidics 21
     2.2.3 GCI vs other label-free biosensor technologies 24
2.3 Overview of applications 26
     2.3.1 Kinetics and affinity analysis 26
     2.3.2 Molecular screening 29
     2.3.3 Binding specificity 31
     2.3.4 Concentration analysis 32
3. Experimental design 34
3.1 Sensor Surfaces 35
     3.1.1 Chip type selection 35
     3.1.2 Chip conditioning 40
     3.1.3 Immobilization strategies 41
     3.1.4 Surface regeneration 51
     3.1.5 Ligand regeneration 53
3.2 Ligand immobilization 56
     3.2.1 Ligand density 56
     3.2.2 Ligand activity 59
     3.2.3 Mass transport limitation 60
     3.2.4 Ligand avidity 61
     3.2.5 Non-specific binding 62
3.3 Buffer 63
     3.3.1 General considerations 63
     3.3.2 Buffer composition 63
     3.3.3 Buffer/sample matching 67
3.4 Referencing and correction 69
     3.4.1 Referencing 69
     3.4.2 Double referencing 69
     3.4.3 DMSO correction (solvent correction) 70
3.5 Assay format 75
     3.5.1 Kinetics and affinity analysis 75
     3.5.2 Molecular screening 84
     3.5.3 Binding specificity and competition experiments 85
     3.5.4 Concentration analysis 87
     3.5.5 Sample recovery for analyte binder identification 91
4. Data analysis 97
4.1 Data adjustment 98
     4.1.1 Signal jumps 99
     4.1.2 Offsets 99
     4.1.3 DMSO correction (solvent correction) 100
     4.1.4 Blanks 101
4.2 Data evaluation 102
     4.2.1 Evaluation output and quality 103
     4.2.2 Settings and advanced menu 105
     4.2.3 Model selection 109
     4.2.4 Equilibrium analysis 116
4.3 waveRAPID 117
4.4. Screen 119
     4.4.1 Settings 119
     4.4.2 Plot data series 119
     4.4.3 Ranking/hits 120
     4.4.4 Off-rate screens 120
5. Troubleshooting 122
5.1 Leakage 123
5.2 Device state 123
5.3 Distorted signa 124
     5.3.1 Noisy signal 124
     5.3.2 Unstable/low amplitudes 125
     5.3.3 Signal spikes/phase losses 126
5.4 Ligand immobilization 127
     5.4.1 Low binding 127
     5.4.2 Low ligand activity 128
5.5 Kinetics 128
     5.5.1 No saturation/over-stoichiometric signal 128
     5.5.2 No dissociation signal/ linear association 129
     5.5.3 waveRAPID 129
6. References 130

All WAVEsystem products have been designed with user experience in mind. At the same time, they are highly sophisticated bioanalytical instruments requiring skilled user input. 

To help you get the most from your product, we have combined the wealth of our internal knowledge and experience in one practical reference manual. Our experts guide you through the main principles underlying the technologies, techniques and applications. We provide information, advice and tips on experimental design and data analysis. A separate troubleshooting section covers the situations and questions encountered most often by our customer support teams.

Whether you read it from cover to cover, refer to it for specific questions, or keep it as a handy reminder of equations and protocols, we hope that this manual will support you in your work and contribute to results that accelerate discovery.

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[Creoptix kinetics guide.jpg] Creoptix kinetics guide.jpg

What's in the guide?

Content Page
1. Welcome 5
2. Introduction 7
2.1 Introduction to label-free interaction analysis 8
     2.1.1 Definitions of label-free terminology 12
     2.1.2 Principles of label-free analysis 16
2.2 The Creoptix WAVEsystem 19
     2.2.1 GCI technology 19
     2.2.3 Non-clog microfluidics 21
     2.2.3 GCI vs other label-free biosensor technologies 24
2.3 Overview of applications 26
     2.3.1 Kinetics and affinity analysis 26
     2.3.2 Molecular screening 29
     2.3.3 Binding specificity 31
     2.3.4 Concentration analysis 32
3. Experimental design 34
3.1 Sensor Surfaces 35
     3.1.1 Chip type selection 35
     3.1.2 Chip conditioning 40
     3.1.3 Immobilization strategies 41
     3.1.4 Surface regeneration 51
     3.1.5 Ligand regeneration 53
3.2 Ligand immobilization 56
     3.2.1 Ligand density 56
     3.2.2 Ligand activity 59
     3.2.3 Mass transport limitation 60
     3.2.4 Ligand avidity 61
     3.2.5 Non-specific binding 62
3.3 Buffer 63
     3.3.1 General considerations 63
     3.3.2 Buffer composition 63
     3.3.3 Buffer/sample matching 67
3.4 Referencing and correction 69
     3.4.1 Referencing 69
     3.4.2 Double referencing 69
     3.4.3 DMSO correction (solvent correction) 70
3.5 Assay format 75
     3.5.1 Kinetics and affinity analysis 75
     3.5.2 Molecular screening 84
     3.5.3 Binding specificity and competition experiments 85
     3.5.4 Concentration analysis 87
     3.5.5 Sample recovery for analyte binder identification 91
4. Data analysis 97
4.1 Data adjustment 98
     4.1.1 Signal jumps 99
     4.1.2 Offsets 99
     4.1.3 DMSO correction (solvent correction) 100
     4.1.4 Blanks 101
4.2 Data evaluation 102
     4.2.1 Evaluation output and quality 103
     4.2.2 Settings and advanced menu 105
     4.2.3 Model selection 109
     4.2.4 Equilibrium analysis 116
4.3 waveRAPID 117
4.4. Screen 119
     4.4.1 Settings 119
     4.4.2 Plot data series 119
     4.4.3 Ranking/hits 120
     4.4.4 Off-rate screens 120
5. Troubleshooting 122
5.1 Leakage 123
5.2 Device state 123
5.3 Distorted signa 124
     5.3.1 Noisy signal 124
     5.3.2 Unstable/low amplitudes 125
     5.3.3 Signal spikes/phase losses 126
5.4 Ligand immobilization 127
     5.4.1 Low binding 127
     5.4.2 Low ligand activity 128
5.5 Kinetics 128
     5.5.1 No saturation/over-stoichiometric signal 128
     5.5.2 No dissociation signal/ linear association 129
     5.5.3 waveRAPID 129
6. References 130

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