Drug selection for breast cancer therapy using antibody-based arrays
AssigneePrometheus Laboratories Inc.
InventorsSharat Singh, Jeanne Harvey, Phillip Kim, Xinjun Liu, Limin Liu, Robert Barham, Bruce Neri
Life of this patent
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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.
Claims71
A method for selecting a suitable anticancer drug for the treatment of a breast tumor, the method comprising:
- (a) isolating cells of a breast tumor after administration of an anticancer drug, or prior to incubation with an anticancer drug;
- (b) lysing the isolated 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
- (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.
A method for identifying the response of a breast tumor to treatment with an anticancer drug, the method comprising:
- (a) isolating cells of a breast tumor after administration of an anticancer drug, or prior to incubation with an anticancer drug;
- (b) lysing the isolated 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
- (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.
-80. (canceled)
A method for predicting the response of a subject having a breast tumor to treatment with an anticancer drug, the method comprising:
- (a) isolating cells of a breast tumor after administration of an anticancer drug, or prior to incubation with an anticancer drug;
- (b) lysing the isolated 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
- (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.
-99. (canceled)
An array having superior dynamic range comprising a plurality of dilution series of capture antibodies restrained on a solid support, wherein the capture antibodies in each dilution series are specific for one or more analytes corresponding to a component of a signal transduction pathway in a cellular extract.
-104. (canceled)
A method for detecting the presence of a truncated receptor, said method comprising:
- (a) incubating a cellular extract with a plurality of beads specific for an extracellular domain (ECD) binding region of a full-length receptor;
- (b) removing said plurality of beads from said cellular extract, thereby removing said full-length receptor to form a cellular extract devoid of said full-length receptor;
- (c) incubating said cellular extract devoid of said full-length receptor with a plurality of capture antibodies, wherein said plurality of capture antibodies is specific for an intracellular domain (ICD) binding region of said truncated receptor and wherein said plurality of capture antibodies is restrained on a solid support to foam a plurality of captured truncated receptors;
- (d) incubating the plurality of captured truncated receptors with detection antibodies specific for the corresponding truncated receptors to form a plurality of detectable captured truncated receptors;
- (e) incubating the plurality of detectable captured truncated receptors with first and second members of a signal amplification pair to generate an amplified signal;
- and
- (f) detecting an amplified signal generated from the first and second members of the signal amplification pair.
-134. (canceled)
A method for detecting the presence of a truncated receptor, said method comprising:
- (a) incubating a cellular extract with a plurality of beads specific for an extracellular domain (ECD) binding region of a full-length receptor;
- (b) removing said plurality of beads from said cellular extract, thereby removing said full-length receptor to form a cellular extract devoid of said full-length receptor;
- (c) incubating said cellular extract devoid of said full-length receptor with a plurality of capture antibodies, wherein said plurality of capture antibodies is specific for an intracellular domain (ICD) binding region of said truncated receptor and wherein said plurality of capture antibodies is restrained on a solid support to form a plurality of captured truncated receptors;
- (d) incubating the plurality of captured truncated receptors with detection antibodies comprising a plurality of activation state-independent antibodies and a plurality of activation state-dependent antibodies specific for the corresponding truncated receptors to form a plurality of detectable captured truncated receptors,
- wherein the activation state-independent antibodies are labeled with a facilitating moiety, the activation state-dependent antibodies are labeled with a first member of a signal amplification pair, and the facilitating moiety generates an oxidizing agent which channels to and reacts with the first member of the signal amplification pair;
- (e) incubating the plurality of detectable captured truncated receptors with a second member of the signal amplification pair to generate an amplified signal;
- and
- (f) detecting the amplified signal generated from the first and second members of the signal amplification pair.
-168. (canceled)
Dependencies are inferred from the claim text.