Fluidity One-W
Investigate protein interactions in solution
Fluidity One systems offer a novel approach to determine affinity (KD), size, concentration, and stoichiometric information when a labeled protein interacts with DNA, lipids, antigens or other proteins
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Analyze protein interactions in aqueous buffer solutions
Eliminate risk of artefacts
No surface attachment -
Determine affinity KD and concentration
Control for off-target binding and
false positives -
Verify the identity of a complex
Confirm interaction partners via absolute size (hydrodynamic radius, Rh) measurements
Infer stoichiometry of interaction
Data in agreement with reported literature
PD-1/PD-L1 interaction – critical in control of autoimmune response
A comparison of binding affinity values for PD-1/PD-L1 from a range of biophysical techniques
| KD (µM) | Technique | Reference |
| 4.0 | Fluidity One-W | |
| 2.2 | ITC | Cheng et al., JBC (2013) |
| 3.9 | SPR equilibrium | Zhang et al., Immunity (2004) |
| 0.7 | SPR equilibrium | Latchman et al., Nat. Imm. (2001) |
| 7.2 | MST | Magnez et al., Sci. Rep. (2017) |
Protein-DNA interactions
KD of aptamer-protein interactions
- No calibration or specialist preparation required
- KD successfully calculated for each aptamer binding
- KD values in agreement with reported literature values
Reveal changes in molecular conformation
Follow changes in hydrodynamic radius in solution: Protein A:IgG interaction
- Determine size of SpA:IgG complex in solution
- Microfluidic diffusion sizing confirmed SpA:IgG 1:3
- Experimental data in agreement with hypothetical Rh values from molecular modelling (PyMOL*)
Take a closer look
Fluidity One-W highlights
- Generate data in close to physiologically-relevant conditions
- Reduce or eliminate need for time-consuming purification
- Determine KD and concentration directly in solution
- Minimize sample consumption – typically 60 – 100 μL per KD
- Reveal changes in molecular conformation
Fluidity One-W specifications
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System
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Application
Determine size, KD and stoichiometry in aqueous buffer systems
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Technology
Microfluidic Diffusional Sizing (MDS)
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Interaction analysis
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Run time
Typically 8 – 14 min per datapoint
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Size range: hydrodynamic radius, Rh
1 – 20 nm
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Accuracy of size determination
± 10%
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Reproducibility of size determination
CV < 10%
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Working range molecular weight
1.4 kDa – 14 MDa
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Sensitivity range (labeled HSA in PBS)
1 nM – 1 μM Alexa Fluor™ 488
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Typical sample consumption to determine protein KD
60 – 100 μL
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Sample volume per datapoint
5 μL
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Compatibility
Compatible with aqueous buffer systems
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Fluorescent labels
Alexa Fluor™ 488 and equivalents | GFP and FITC | Fluidiphore labeling kit (fluidiphore rapid amine 503)
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Data export
USB Mass Storage Device | Fluidity Cloud
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Exported data file formats
CSV and JSON formats
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Data output from Fluidity Cloud
Result tables, binding curves and affinity (KD), size (Rh) of complex and labeled species
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Consumables
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Consumables
Kits containing chips and cartridges sufficient for 96 or 288 datapoints
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Specifications
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Temperature control
Ambient
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Operating environment
5 °C to 40 °C
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Power requirements
100 – 240 V AC, 50 – 60 Hz
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Safety and EMC standards
Designed to comply with all relevant safety and EMC standards
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Dimensions
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Dimensions (D x W x H; mm)
400 x 400 x 430
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Weight (kg)
15 kg
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Publications and resources
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Fluidic Analytics is changing how proteins are characterised
We're doing this with novel, cutting edge technology and a fantastic, committed team
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