Patent NZ-587420-A (NZ)
Title: Drug selection for breast cancer therapy using antibody-based arrays
Inventor: SINGH SHARAT
IPC: G01N33/50
Filed: 2009-02-24
Abstract: method for selecting a suitable anticancer d 1 rug for the treatment of a breast tumor, the method comprising: (a) lysing cells of a breast tumor isolated after administration of an anticancer drug, or prior to incubation with an anticancer drug to produce a cellular extract; (b) detecting an activation state of one or more analytes in the cellular extract using an assay comprising a plurality of dilution series of capture antibodies specific for the one or more analytes, wherein the capture antibodies are restrained on a solid support; and wherein the assay comprises: (i) incubating the cellular extract with the plurality of dilution series of capture antibodies to form a plurality of captured analytes; (ii) incubating the plurality of captured analytes with detection antibodies comprising a plurality of activation state-independent antibodies and a plurality of activation state-dependent antibodies specific for the corresponding analytes to form a plurality of detectable captured analytes, wherein the activation state-independent antibodies are labeled with glucose oxidase, wherein the glucose oxidase and the activation state20 independent antibodies are conjugated to a sulfhydryl-activated dextran molecule, wherein the activation state-dependent antibodies are labeled with a first member of a signal amplification pair, and wherein the glucose oxidase generates an oxidizing agent which channels to and reacts with the first member of the signal amplification pair; (iii) incubating the plurality of detectable captured analytes with a second member of the signal amplification pair to generate an amplified signal; and (iv) detecting the amplified signal generated from the first and second members of the signal amplification pair; and (c) determining whether the anticancer drug is suitable or unsuitable for the Provided is a method for selecting a suitable breast cancer drug or identifying a response to treatment by a breast cancer drug, the method comprising: (a) lysing cells of a breast tumor isolated after administration of an anticancer drug, or prior to incubation with an anticancer drug to produce a cellular extract; (b) detecting an activation state of one or more analytes in the cellular extract using an assay comprising a plurality of dilution series of capture antibodies specific for the one or more analytes, wherein the capture antibodies are restrained on a solid support; and wherein the assay comprises: (i) incubating the cellular extract with the plurality of dilution series of capture antibodies to form a plurality of captured analytes; (ii) incubating the plurality of captured analytes with detection antibodies comprising a plurality of activation state-independent antibodies and a plurality of activation state-dependent antibodies specific for the corresponding analytes to form a plurality of detectable captured analytes, wherein the activation state-independent antibodies are labelled with glucose oxidase, wherein the glucose oxidase and the activation state independent antibodies are conjugated to a sulfhydryl-activated dextran molecule, wherein the activation state-dependent antibodies are labelled with a first member of a signal amplification pair, and wherein the glucose oxidase generates an oxidizing agent which channels to and reacts with the first member of the signal amplification pair; (iii) incubating the plurality of detectable captured analytes with a second member of the signal amplification pair to generate an amplified signal; and (iv) detecting the amplified signal generated from the first and second members of the signal amplification pair; and (c) determining whether the anticancer drug is suitable or unsuitable for the treatment of the breast tumor by comparing the activation state detected for the one or more analytes with a reference activation profile generated in the absence of the anticancer drug.