Biological encoding of large numbers of cells
AssigneePrimity Bio, Inc.
InventorsPeter Oliver Krutzik, Thomas Scott Wehrman
Life of this patent
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Abstract
Mixtures of cell types can be analyzed by having at least two signal markers, with at least one at three different levels to provide a barcode for each cell type. The mixture of cells may be subjected to a common candidate moiety and the effect of the moiety on the cells determined along with identification of the cell by the barcode. Conveniently, surface marker proteins and labeled antibodies can be used to create the barcode and the cells analyzed with flow cytometry.
Claims24
A method of distinguishing between cell types in a sample comprising a plurality of different cell types comprising:
- a) labeling at least two different signal markers expressed by at least one cell type in the sample with distinguishably labeled binding proteins, wherein the at least two different signal markers are exogenous cell surface proteins, wherein said exogenous cell surface proteins are constitutively expressed from one or more genetic constructs, and wherein at least one of said signal markers is expressed at least at three different amounts in said plurality of different cell types;
- b) detecting from at least one cell type of said plurality of different cell types the at least two different, distinguishably labeled signal markers;
- c) measuring at least one of said labeled signal markers of step b) at three different intensities corresponding to said at least three different amounts in the plurality of different cell types, wherein the combination of the intensity and type of signal markers in a single cell type provides a unique barcode;
- and
- d) distinguishing at least one cell type of said plurality of different cell types based on the unique barcode provided in c).
A method of distinguishing between cell types in a sample comprising at least 6 different cell types comprising:
- a) labeling at least two different signal markers expressed by at least one cell type in the sample with fluorescently labeled antibodies, wherein the at least two different signal markers are exogenous cell surface proteins, wherein said at least two signal markers are distinguishably labeled, and wherein said exogenous cell surface proteins are constitutively expressed from one or more genetic constructs;
- b) detecting from at least one cell type of said 6 different cell types the at least two different, distinguishably labeled signal markers by flow cytometry, wherein at least one of said signal markers is expressed at least at three different amounts in said at least 6 different cell types;
- c) measuring at least one of said labeled signal markers of step b) at three different intensities, wherein the combination of the intensity and type of signal markers in a single cell type provides a unique barcode;
- and
- d) distinguishing at least one cell type of said plurality of different cell types based on the unique barcode provided in c).
A mixture of cells comprising at least 6 different cell types, wherein:
- a) each cell type comprises at least two genetic constructs which express cell surface proteins exogenous to said cell types;
- b) at least on of said cell surface proteins is configured to be expressed at least at three different amounts with a different amount in each of said cell types, wherein the expression of the at least one cell surface protein is detectable at least at three different intensities;
- and
- c) said cell types can be distinguished by detecting binding of labeled antibodies to each of said cell surface proteins of each cell, wherein each cell surface protein binds to a distinguishably labeled antibody.
The mixture of 23, wherein said labeled antibodies are bound to each of said cell surface proteins.
Dependencies are inferred from the claim text.