US 2006/0040407 A1 · Alleviation of non-specific binding in microarray assays

Abstract

A post-incubation treatment is employed to effectively remove targets, such as proteins/protein complexes, or other label-bearing moieties that may non-specifically bind to a microarray substrate during a binding assay. Following incubation, a one-step wash is carried out with a liquid containing digester, e.g., a digestive enzyme (protease) or lysosome, which is effective to remove non-specifically bound targets or at least labeled portions of such targets from the substrate. Proteases are bound to or coated onto large molecules or onto solid particles of such a size such that they are prohibited from entering the porous surfaces of 3-D hydrogel microspots and are unable to reach and digest labeled target-probe complexes that are disposed within such porous hydrogel microspots. Digested segments of such protein which contain labels (or of essentially the entire protein) are carried away in the wash liquid and thus are not present to create background noise during imaging.

Claims20

Dependent claims

A method for the determination of the presence of one or more targets in a sample being analyzed, which method comprises:

providing a substrate having an upper surface,
attaching a plurality of 3-dimensional porous microspots of hydrogel to said surface,
providing different capture agents within the pores of various of said plurality of microspots to create a microarray on said surface,
contacting said microarray with a solution containing targets of a size that allows binding with said capture agents within said porous hydrogel microspots,
washing said microarray following said contacting using a liquid containing a digester which removes non-specifically bound targets or at least labeled portions thereof from said surface without affecting targets specifically bound to capture agents within said porous hydrogel microspots,
associating labels with said targets either before or after said washing if said targets do not comprise labels at the time of said binding, and
measuring said washed microarray to detect signals from labeled targets bound within particular microspots.

4 more elements

Dependencies are inferred from the claim text.

Alleviation of non-specific binding in microarray assays

Life of this patent

Alleviation of non-specific binding in microarray assays

Life of this patent

Abstract

Claims20