Biocept, Inc.

Yehudit Falcovitz-Gerassi

Pavel Tsinberg

Soonkap Hahn

Pat Roycroft

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.

Alleviation of non-specific binding in microarray assays

Microwell biochips () are formed from a thin flat plate () of polymeric material having a plurality of regularly spaced holes () that extend completely therethrough and create microwells. The lower end of each hole is closed by a microporous, hydrophobic, polymeric membrane () laminated to the undersurface of the plate which retains an aqueous test solution in the wells until a vacuum is applied to the undersurface thereof to effect draining of the solution and of any wash solution that might be subsequently added. A spot of polymerizing isocyanate-functional hydrogel is applied generally centrally to the porous membrane surface at the bottom of each well in a manner so as to cover only a minor portion of the surface and out of contact with the well sidewalls, thus leaving substantial surface area through which drainage can be readily effected. Biological capture agents are associated with the polymerizing hydrogel so as to become immobilized as a part thereof.

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San Diego, CA, United States of America

Yehudit Falcovitz-Gerassi