RESOURCES

Name of Method: SPR

Result expected: Information on specificity, kinetics, affinities, drug screening (inhibition of complex binding)

K_D range:
Can characterize interactions with K_D more than 10^-10 M and less than 10^-2 M

MW range: Minimum ~1 kDa detectable for analyte

Purity: Crystallisation grade (95%) for binding/kinetic data, lower for drug screening

Labeling-Tagging: Should any or all of reactants be labeled or tagged and how ?
Yes - depending on chips used - choice of His-tag (less-predictable), biotin tag (best). Tag not absolutely necessary - can use amine coupling but it's less predictable.

Accuracy: (Da) N/A

Quantity required: For a complex AB, assuming that A is the 'receptor' and B the 'ligand' how much protein is needed to characterize the complex fully (e.g. obtain K_D or get MW) ?
A total of 100-200 µl of A, at a concentration of 5 x10^-4 - 1 mg/ml
A total of 200-300 µl of B at a concentration of 10 mg/ml

HTP: Is the method HTP (can it use 96-well sample plates in an automated fashion)?
Yes - at NKI

Speed: Will the user have a quantitative answer immediately after collecting the data?
Not for affinity/kinetic data but yes for drug screening - answer after evaluation

Cost: (approximate cost of consumables per ONE experiment)
Approx €150-300 per experiment

Access: Is an instrument available (name all SPINE2 sites you know)?
Oxford, Amsterdam, Strasbourg, Florence

User expertise: Can the method be provided as a service (measurement and data analysis done by local staff) or only access and assistance on a collaborative basis?
Available on a collaborative basis

Special requirements: Any special sample needs?
Pure sample - possibly need to biotinylate immobilized ligand for best results; filtered through 0.2?M