Labeled cell sets for use as functional controls in rare cell detection assays
AssigneeImmunivest Corporation
InventorsGalla Chandra Rao, Herman Rutner
Life of this patent
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Abstract
Methods and kits for used as an internal or external control in rare cell analysis are disclosed. A plurality of fluorescently distinct sets of cells is used to define a range to assess the isolation of rare target cells from a sample. Thus, a known number of cells, expressing the surface and intracellular antigens present in the targeted rare cells, are stabilized and modified in such a way that they can be discriminated from the targeted rare cells. In addition, these cells are separated into at least two sets based upon the number. These sets are detectably distinct from each other and provide an upper and lower indication of the detection ability of the rare cell assay.
Claims22
An improved method for detecting and enumerating rare cells in a mixed cell population by obtaining a blood specimen from a test subject, said specimen comprising a mixed cell population suspected of containing said rare cells, which method comprises preparing an immunomagnetic sample wherein said blood specimen is mixed with magnetic particles coupled to a ligand which reacts specifically with a determinant of said rare cells, to the substantial exclusion of other sample components, contacting said immunomagnetic sample with at least one reagent which labels said determinant of said rare cells, and analyzing said labeled rare cells to determine the presence and number of any said rare cells in said immunomagnetic sample, wherein the greater the number of said rare cells present in said sample, the greater the severity of a disease state, and wherein the improvement comprises the addition of two or more fluorescently distinct sets of stabilized cell populations, permeabilized, for use as a range of control cells in said method, and wherein the membranes of said control cells are detectably labeled, and wherein said control cells contain stabilized cellular components and antigenic moieties of said control cells stabilized for a period up to six months by exposure to fixative.
An improved kit for screening a patient sample for the presence of circulating tumor cells, said kit having coated magnetic nanoparticles with a magnetic core material, a protein base coating material, and an anti-EpCAM coupled, directly or indirectly, to said base coating material, at least one antibody having binding specificity for a cancer cell determinant, and cell specific dye for excluding sample components other than said tumor cells from analysis, wherein the improvement comprises the addition of two distinct sets of stabilized cell populations, permeabilized, for use as control cells in said kit, said distinct sets of stabilized control cell populations having determinants in common with said rare cells, wherein said membranes of said control cells are detectably labeled, and cellular components and antigenic moieties of said control cells have been stabilized up to six months, said stabilized control cells being suspended in a buoyant density medium
Dependencies are inferred from the claim text.