Methods and reagents for the rapid and efficient isolation of circulating cancer cells
OwnerLeon W.M.M. Terstappen, Galla Chandra Rao +4inventors
InventorsLeon W.M.M. Terstappen, Galla Chandra Rao, Shawn Mark O'Hara, Paul A. Liberti, Steven Gross, Gerald Doyle
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Abstract
Methods and compositions are provided for detecting circulating tumor cells and assessing said cells for alterations in tumor-diathesis associated molecules.
Claims82
A method for assessing a patient for the presence of a malignancy, comprising:
- a) obtaining a biological specimen from a patient, said specimen comprising a mixed cell population suspected of containing hematopoietic and non-hematopoietic malignant cells;
- b) preparing a sample wherein said biological specimen is mixed with a detectably labeled ligand which reacts specifically with the malignant cells, to the substantial exclusion of other sample components;
- c) contacting said sample with at least one reagent which also specifically labels said malignant cells;
- d) analyzing said sample to determine the presence and number of labeled cells, detection of said cells indicating the presence of malignancy, the greater the number of cells present, the greater the severity of the malignancy;
- wherein the method further comprises assessment of said labeled cells for alterations in at least one tumor diathesis-associated molecule.
A method for assessing a patient for the presence of a non-hematopoietic malignancy, comprising:
- a) obtaining a biological specimen from a patient, said specimen comprising a mixed cell population comprising hematopoietic and non-hematopoietic malignant cells;
- b) preparing an immunomagnetic sample wherein said biological specimen is mixed with magnetic particles coupled to a ligand which reacts specifically with the malignant cells, to the substantial exclusion of other sample components;
- c) separating said magnetic particle containing malignant cells from non-magnetic particle hematopoietic cells, and
- d) analyzing said magnetic particle-containing cells to determine the presence and number of any malignant cells in said sample, detection of said cells indicating the presence of malignancy, the greater the number of cells present, the greater the severity of the malignancy;
- wherein the method further comprises assessment of said magnetic particle containing malignant cells for alterations in tumor diathesis-associated cellular molecules.
A method for determining alterations in tumor diathesis associated molecules as a means to predict efficacy of therapy, comprising:
- a) obtaining a sample from a patient;
- b) isolating and enumerating circulating malignant cells from said sample if present, said method further comprising;
- and
- c) determining the number of at least one predetermined tumor diathesis associated molecule on individual cells present in said sample as a means to predict efficacy of therapy.
A method for determining alterations in tumor diathesis associated molecules as a means to assess appropriate dosage for therapy, comprising:
- a) obtaining a sample from a patient;
- b) isolating and enumerating circulating malignant cells from said sample if present, said method further comprising;
- and
- c) determining the number of at least one predetermined tumor diathesis associated molecule on individual cells present in said sample as a means to assess appropriate dosage for therapy.
A method for determining alterations in tumor diathesis associated molecules as a means to monitor efficacy of therapy, comprising:
- a) obtaining a sample from a patient;
- b) isolating and enumerating circulating malignant cells from said sample if present, said method further comprising;
- and
- c) determining the number of at least one predetermined tumor diathesis associated molecule on individual cells present in said sample as a means to monitor efficacy of therapy.
A method for determining alterations in tumor diathesis associated molecules as a means to assess cancer progression comprising:
- a) obtaining a sample from a patient;
- b) isolating and enumerating circulating malignant cells from said sample if present, said method further comprising;
- and
- c) determining whether said cells contain a predetermined tumor diathesis associated molecule associated with a poor prognosis as a means to assess cancer progression.
A method for performing a whole body biopsy on a patient, comprising:
- a) obtaining a blood sample from a patient;
- b) isolating and enumerating circulating non-hematopoietic malignant cells from said sample if present, said method further comprising;
- and
- c) analyzing said cells for the presence and number of a panel of predetermined tumor diathesis associated molecules.
A method for identifying alterations in a circulating tumor cells relative to cells present in a tumor mass in situ, comprising:
- a) obtaining a biopsy specimen of said tumor mass from patient;
- b) isolating circulating tumor cells from said patient, if present;
- c) contacting said specimen and said isolated circulating tumor cells with a duplicate panel of agents which detect a plurality of tumor diathesis associated molecules;
- d) detecting any tumor diathesis associated molecules present in said circulating tumor cells and in said specimen;
- and
- e) determining whether said tumor diathesis associated molecules are altered in said circulating tumor cell relative to said biopsy specimen.
A test kit for screening a patient sample for the presence of a non-hematopoietic malignant cells comprising:
- a) coated magnetic nanoparticles comprising a magnetic core material, a protein base coating material, and an antibody that binds specifically to a first characteristic determinant of said malignant cell, said antibody being coupled, directly or indirectly, to said base coating material;
- b) at least one antibody having binding specificity for a second characteristic determinant of said malignant cell;
- c) a cell specific dye for excluding sample components other than said malignant cells from analysis;
- d) a device selected from the group consisting of a Cell Spotter® cartridge or a Cell Tracks cartridge;
- and
- e) at least one detectably labeled agent having binding affinity for a tumor diathesis associated molecule.
Dependencies are inferred from the claim text.