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