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

•Reliable affinity measurement

•Good for screening protein-protein interaction

•Not limited by surface chemistry

•Relying on fluorescent labelling

•Faster learning curve

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

•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)

Platform of choice:		SPR	MST	BLI	FIDA
Sample type:	crude extract/serum	✓(suboptimal)		✓	✓ (possibility of aggregates depending on interactions)
Sample type:	Purified protein/small molecule	✓	✓	✓	✓
Analyte	Purified protein	✓	✓	✓	✓
Analyte	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		✓	✓	✓
Is buffer exchange a limitation for your protein?	No	✓
Is surface chemistry a limitation for your protein?	Yes		✓		✓
Is surface chemistry a limitation for your protein?	No	✓		✓

If you need further help please contact:

BioSRLs@warwick.ac.uk