Feb 24, 2025
Kinetic assay with waveRAPID grating-coupled interferometry waveCreoptix system
- Eda Capkin1,2,3
- 1Diamond Light Source;
- 2Research Complex at Harwell;
- 3ASAP Discovery Consortium
- ASAP Discovery
Protocol Citation: Eda Capkin 2025. Kinetic assay with waveRAPID grating-coupled interferometry waveCreoptix system. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk87xwl5r/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 22, 2024
Last Modified: February 24, 2025
Protocol Integer ID: 110525
Keywords: Molecular interactions, Label-free detection, Grated-coupled interferometry, Automated sampling, coupled interferometry wavecreoptix system, interferometry wavecreoptix system this protocol, binding assay, creoptix waverapid grating, molecular interaction measurement, throughput molecular interaction measurement, waverapid grating, interferometry system, coupled interferometry system
Funders Acknowledgements:
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
Grant ID: Grant ID: U19AI171399
Disclaimer
The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Abstract
This protocol outlines the setup of kinetic binding assays using the Creoptix waveRAPID grating-coupled interferometry system. The method covers automated sample handling in 96- or 384-well formats, including DMSO corrections, control samples, and buffer blanks, enabling high-throughput molecular interaction measurements.
Materials
10x HBS-P:HBS-P Buffer 10×CytivaCatalog #BR100671
DMSO: Pierce™ Dimethylsulfoxide (DMSO), Sequencing GradeThermo ScientificCatalog #10127403
Echo 384 well plate: Echo® Qualified 384-Well Polypropylene Microplate 2.0Beckman CoulterCatalog #001-14555
Creoptix 384 well plate:240 µl 384 Deepwell Plate, Square Wells StarLabCatalog #E2384-2400
Creoptix 96 well plate: PlateOne 96 well Conical Bottom (V) Plates, max. Working Volume 320 µlStarLabCatalog #S1833-9600
Troubleshooting
Safety warnings
Do not set the Flow Cell Temperature below 10 and/or 15°C. Lower temperature causes to the condensation in the chip.
Before start
Once the concentration value is specified in the method, it can not be changed in the evaluation part.
RAPID kinetics assay
5m
Variables that are for RAPID kinetic assay steps indicated the table.
| A | B | |
| Variable | Abbrevation for variable | |
| Running buffer+DMSO% | [X] | |
| Sample concentration | [Y] | |
| Association and dissociation duration | [Z] | |
| Flow rate | [F] |
These specifics will differ for each protein target.
Build the method in the Measure tab/ RAPID kinetics.
In the Wizard choose the correct sample container type for the experiment: 48 vials, 96 or 384 well plate.
Enter the details of samples including sample name, concentration, and molecular weight.
Enter the configuration details for the method. Specify flow rate [F] and association and dissociation duration [Z].
In the Reagent Rack section, choose 10 startup cycles.
Select Blanks: 1 blank after every 5 samples.
Choose DMSO correction ( 0.5%DMSO added to running buffer ) as DMSO calibration solution at the beginning, end + every 10 th sample.
Enter a control sample and indicate its molecular weight and repeats depending on the sample size.
(i.e. For 96 samples, choose 1 sample injection after 50 th sample.)
Select DMSO wash: 1 DMSO wash (50 % volume DMSO) after every 5 samples.
Click create series.
Check the required buffer volume from cycles tab within the configuration. Prepare running
buffer including DMSO [X] about 200 ml more than calculated buffer volume.
Degas the running buffer with DMSO% for 2 minutes.
2m
Sample preparation
Transfer samples from 100 millimolar (mM) stocks 1536LDV to 96 or 384 well plate and add running buffer +DMSO% [X] to make final µM sample concentration [Y].
Blank, DMSO calibration solution, and Control preparation
Go to the Autosampler section within the configuration and prepare other solution (running buffer as blank and start-ups, DMSO calibration solution and control sample).
Place the samples according to the series in autosampler.
Run 2x Priming and Chip Prime steps in Device/Chip Prime
Go to the Series/Run Measurement.
Protocol references
Kartal Ö, Andres F, Lai MP, Nehme R, Cottier K. waveRAPID-A Robust Assay for High-Throughput Kinetic Screens with the Creoptix WAVEsystem. SLAS Discov. 2021 Sep;26(8):995-1003. doi: 10.1177/24725552211013827. Epub 2021 May 28. PMID: 34049465.