Biomarker detection in circulating cells
First Claim
1. A method of detecting one or more protein-protein complexes of a rare cell type in a sample containing a mixed population of cells such that each protein-protein complex has a first protein and a second protein, the method comprising:
- immunomagnetically isolating from the sample a subpopulation of cells containing the rare cell type with one or more antibody compositions, wherein each antibody composition is specific for a cell surface antigen of the rare cell type and wherein each antibody composition is attached to a magnetic particle;
combining with the subpopulation of cells(a) one or more different first binding compounds specific for each first protein of each of the one or more protein-protein complexes of the rare cell type, wherein each first binding compound has one or more molecular tags attached thereto via a cleavable linkage, the one or more molecular tags of each different binding compound having a distinct separation characteristic so that the one or more molecular tags of each different binding compound form distinct peaks in a separation profile upon separation; and
(b) one or more different second binding compounds specific for each second protein of each of the one or more protein-protein complexes of the rare cell type, wherein each second binding compound is conjugated to a cleaving-inducing moiety having an effective proximity;
wherein each of the first binding compounds specifically binds to each first protein and each of the second binding compounds specifically binds to each second protein, and wherein the one or more molecular tags of each first binding compound are released; and
separating and identifying the released by the cleaving-inducing moiety when in effective proximity to said one or more molecular tags molecular tags to detect the one or more protein-protein complexes in the sample.
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Abstract
A method is provided for detecting biomarkers in rare circulating cells by forming an enriched population of cells immunomagnetically followed by biomarker detection using binding compounds having releasable molecular tags. Preferably, biomarkers comprising one or more protein-protein complexes are detected in circulating cancer cells metastasized from a solid tumor. In the presence of biomarkers, the molecular tags of the binding compounds are released and separated from the assay mixture for detection and/or quantification.
152 Citations
11 Claims
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1. A method of detecting one or more protein-protein complexes of a rare cell type in a sample containing a mixed population of cells such that each protein-protein complex has a first protein and a second protein, the method comprising:
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immunomagnetically isolating from the sample a subpopulation of cells containing the rare cell type with one or more antibody compositions, wherein each antibody composition is specific for a cell surface antigen of the rare cell type and wherein each antibody composition is attached to a magnetic particle; combining with the subpopulation of cells (a) one or more different first binding compounds specific for each first protein of each of the one or more protein-protein complexes of the rare cell type, wherein each first binding compound has one or more molecular tags attached thereto via a cleavable linkage, the one or more molecular tags of each different binding compound having a distinct separation characteristic so that the one or more molecular tags of each different binding compound form distinct peaks in a separation profile upon separation; and (b) one or more different second binding compounds specific for each second protein of each of the one or more protein-protein complexes of the rare cell type, wherein each second binding compound is conjugated to a cleaving-inducing moiety having an effective proximity; wherein each of the first binding compounds specifically binds to each first protein and each of the second binding compounds specifically binds to each second protein, and wherein the one or more molecular tags of each first binding compound are released; and separating and identifying the released by the cleaving-inducing moiety when in effective proximity to said one or more molecular tags molecular tags to detect the one or more protein-protein complexes in the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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Specification