Surface regeneration of biosensors and characterization of biomolecules associated therewith
AssigneeBiacore AB
InventorsKarl Andersson, Markku Hämäläinen, Magnus Malmqvist, Håkan E. Roos
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Abstract
Surface regeneration of affinity biosensors and characterization of biomolecules associated therewith by multivariate technique employing cocktails of regeneration agents to optimize regeneration of biosensor surface and/or characterize biomolecules associated therewith. Kits and stock solutions for use in the context of this invention, as well as associated computer algorithms are also disclosed.
Claims48
A method for selecting an optimized regeneration solution for the regeneration of a biosensor surface having a surface-bound ligand and an analyte associated with the ligand, comprising:
- (a) sequentially contacting the biosensor surface with each of a plurality of first regeneration cocktails, wherein each of said first regeneration cocktails is an aqueous solution comprising at least one acidic, basic, ionic, organic, detergent or chelating stock solution, and wherein at least one of said first regeneration cocktails comprises a mixture of at least two of said stock solutions;
- (b) measuring the regeneration effect for each of said plurality of first regeneration cocktails to determine which of said plurality of first regeneration cocktails have the highest measured regeneration effect;
- (c) selecting at least two different stock solutions present in said plurality of first regeneration cocktails having the highest measured regeneration effect;
- (d) combining said at least two different stock solutions in various ratios to generate a plurality of second regeneration cocktails;
- (e) sequentially contacting the biosenor surface with each of said plurality of second regeneration cocktails;
- and
- (f) determining the regeneration effect of each of said plurality of second regeneration cocktails and therefrom identifying a second regeneration cocktail as the optimized regeneration solution.
A method for using stored instructions in a computer for selecting an optimized regeneration solution for the regeneration of a biosensor surface having a surface-bound ligand and an analyte associated with the ligand, comprising:
- instructing a device to combine a series of stock solutions in various ratios to generate a plurality of first regeneration cocktails,
- sequentially controlling the device to contact the biosensor surface with each of said plurality of first regeneration cocktails;
- determining the regeneration effect of each of said first regeneration cocktails on the biosensor surface based on measurements received from the device;
- selecting at least two different stock solutions having the highest regeneration effect;
- instructing the device to combine a subset of said at least two different stock solutions in varying ratios to generate a plurality of second regeneration cocktails;
- sequentially controlling the device to contact the biosensor surface with each of said second regeneration cocktails;
- and
- determining the regeneration effect of each of said second regeneration cocktails based on measurements received from the device, and therefrom identifying a second regeneration cocktail as the optimized regeneration solution.
A method for characterizing a ligand or analyte associated with a biosensor surface, comprising the steps of:
- (a) sequentially contacting the biosensor surface having a surface-bound ligand with each of a plurality of characterization solutions;
- (b) introducing the analyte into each of said plurality of characterization solutions so as to interact the analyte with the surface-bound ligand;
- (c) measuring at least one of an association rate, surface-bound analyte concentration, dissociation rate, and regeneration effect of the analyte-ligand interaction for each of said plurality of characterization solutions;
- and
- (d) characterizing the ligand or analyte associated with the biosensor surface based on at least one of the association rate, surface-bound analyte concentration, dissociation rate, and regeneration effect of the analyte-ligand interaction for each of said plurality of characterization solutions.
Dependencies are inferred from the claim text.