Kinetics Guide - Binding kinetics with the WAVEsystem

All Creoptix® products have been designed with user experience in mind. At the same time, they are highly sophisticated devices 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?
1. Welcome5
2. Introduction7
2.1 Introduction to label-free interaction analysis8
     2.1.1 Definitions of label-free terminology12
     2.1.2 Principles of label-free analysis16
2.2 The Creoptix WAVEsystem19
     2.2.1 GCI technology19
     2.2.3 Non-clog microfluidics21
     2.2.3 GCI vs other label-free biosensor technologies24
2.3 Overview of applications26
     2.3.1 Kinetics and affinity analysis26
     2.3.2 Molecular screening29
     2.3.3 Binding specificity31
     2.3.4 Concentration analysis32
3. Experimental design34
3.1 Sensor Surfaces35
     3.1.1 Chip type selection35
     3.1.2 Chip conditioning40
     3.1.3 Immobilization strategies41
     3.1.4 Surface regeneration51
     3.1.5 Ligand regeneration53
3.2 Ligand immobilization56
     3.2.1 Ligand density56
     3.2.2 Ligand activity59
     3.2.3 Mass transport limitation60
     3.2.4 Ligand avidity61
     3.2.5 Non-specific binding62
3.3 Buffer63
     3.3.1 General considerations63
     3.3.2 Buffer composition63
     3.3.3 Buffer/sample matching67
3.4 Referencing and correction69
     3.4.1 Referencing69
     3.4.2 Double referencing69
     3.4.3 DMSO correction (solvent correction)70
3.5 Assay format75
     3.5.1 Kinetics and affinity analysis75
     3.5.2 Molecular screening84
     3.5.3 Binding specificity and competition experiments85
     3.5.4 Concentration analysis87
     3.5.5 Sample recovery for analyte binder identification91
4. Data analysis97
4.1 Data adjustment98
     4.1.1 Signal jumps99
     4.1.2 Offsets99
     4.1.3 DMSO correction (solvent correction)100
     4.1.4 Blanks101
4.2 Data evaluation102
     4.2.1 Evaluation output and quality103
     4.2.2 Settings and advanced menu105
     4.2.3 Model selection109
     4.2.4 Equilibrium analysis116
4.3 waveRAPID117
4.4. Screen119
     4.4.1 Settings119
     4.4.2 Plot data series119
     4.4.3 Ranking/hits120
     4.4.4 Off-rate screens120
5. Troubleshooting122
5.1 Leakage123
5.2 Device state123
5.3 Distorted signa124
     5.3.1 Noisy signal124
     5.3.2 Unstable/low amplitudes125
     5.3.3 Signal spikes/phase losses126
5.4 Ligand immobilization127
     5.4.1 Low binding127
     5.4.2 Low ligand activity128
5.5 Kinetics128
     5.5.1 No saturation/over-stoichiometric signal128
     5.5.2 No dissociation signal/ linear association129
     5.5.3 waveRAPID129
6. References130


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