Molecular interactions techniques selection
SPR - Biacore T200
•Most accurate measurement
•Based on analyte molecular weight (kDa)
•Complete kinetic parameters output
•Limited customisation
•Heavily sensible to buffer used
•Longer run time
MST – Nanotemper Monolith
•Reliable affinity measurement
•Good for screening protein-protein interaction
•Not limited by surface chemistry
•Relying on fluorescent labelling
•Faster learning curve
Fida Neo
- Not limited by surface chemistry
- Label free using 280 nm detector, or with labelling (480 nm and 640 nm detectors)
- Can be used with crude extract and sera as analyte (possibility of aggregates depending on interactions)
- Fast KD screening
BLI – Octet R96
•Kinetic parameters output
•Based on analyte size (nm)
•Faster than SPR
•Not limited by analyte buffer
•Can be used with crude extract and sera as analyte (WHEN APPROPRIATELY DILUTED IN RUNNING BUFFER)
Which is the best platform for my experiment of choice?
| Platform of choice: |
SPR |
MST |
BLI |
FIDA |
|
|---|---|---|---|---|---|
| Sample type: |
crude extract/serum |
✓(suboptimal) |
✓ |
✓ (possibility of aggregates depending on interactions) |
|
| Purified protein/small molecule |
✓ |
✓ |
✓ |
✓ |
|
| Analyte |
Purified protein |
✓ |
✓ |
✓ |
✓ |
| Small molecule |
✓ |
✓ (suboptimal) |
✓ (suboptimal) |
✓ (better to have it as indicator when possible) |
|
| Experiment type: |
Simple KD/Binding screen |
✓ |
✓ |
✓ |
✓ |
| Kinetic parameters |
✓ |
✓ |
✓ |
||
| Target quantification |
✓ |
✓ |
✓ |
||
| Antibody screening |
✓ |
✓ |
✓ |
||
| Competition |
✓ |
✓ (premix only) |
✓ |
✓ (premix only) |
|
| Epitope binning |
✓ |
✓ |
✓ suboptimal, premix only) |
||
| Is buffer exchange a limitation for your protein? |
Yes |
✓ |
✓ |
✓ |
|
| No |
✓ |
||||
| Is surface chemistry a limitation for your protein? |
Yes |
✓ |
✓ |
||
| No |
✓ |
✓ |
|||
If you need further help please contact:
BioSRLs@warwick.ac.uk