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US 2014/0349865 A1Published application
Drug selection for breast cancer therapy using antibody-based arrays
AssigneeNestec S.A.
InventorsSharat Singh, Jeanne Harvey, Phillip Kim, Xinjun Liu, Limin Liu, Robert Barham, Bruce Neri
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Abstract
The present invention provides compositions and methods for detecting the activation states of components of signal transduction pathways in tumor cells. Information on the activation states of components of signal transduction pathways derived from use of the invention can be used for cancer diagnosis, prognosis, and in the design of cancer treatments.
Claims48
A method for selecting a suitable anticancer drug for the treatment of a breast tumor, the method comprising:
- (a) incubating cells isolated from a breast tumor with an anticancer drug in vitro;
- (b) lysing the incubated cells to produce a cellular extract;
- (c) 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 state-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
- (d) 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.
8 more elements
A method for identifying the response of a breast tumor to treatment with an anticancer drug, the method comprising:
- (a) incubating cells isolated from a breast tumor with an anticancer drug in vitro;
- (b) lysing the incubated cells to produce a cellular extract;
- (c) 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 state-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
- (d) identifying the breast tumor as responsive or non-responsive to treatment with the anticancer drug 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.
8 more elements
A method for predicting the response of a subject having a breast tumor to treatment with an anticancer drug, the method comprising:
- (a) incubating cells isolated from a breast tumor with an anticancer drug in vitro;
- (b) lysing the incubated cells to produce a cellular extract;
- (c) 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 state-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
- (d) predicting the likelihood that the subject will respond to treatment with the anticancer drug 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.
8 more elements
Dependencies are inferred from the claim text.