Isolating cells expressing secreted proteins

Patent Number: 6919183

This patent teaches ways of extracting cells from a population based on expression of markers that were either introduced or naturally expressed in the cell. The patent may be useful for pulling out differentiated cells from stem cell heterogenous populations.

A method of detecting and/or isolating a eukaryotic cell that produces a secreted protein of interest (POI), comprising: (a) providing a cell comprising a nucleic acid that encodes a secreted POI and a nucleic acid that encodes a cell surface capture molecule capable of binding the POI, wherein the nucleic acid encoding the secreted POI or the nucleic acid encoding the cell surface capture molecule, or both, are transfected into the cell; (b) culturing the cell under conditions in which the POI and cell surface capture molecule are expressed, and a POI-capture molecule complex is formed intracellularly and displayed on the cell surface; © contacting the cell with a detection molecule, which binds to the POI displayed by the cell; and (d) detecting and/or isolating the cell due to it being bound to the detection molecule.

A method of detecting and isolating a eukaryotic cell that produces a secreted protein of interest (POI), comprising: (a) transfecting the cell with a nucleic acid that encodes a cell surface capture molecule capable of binding the POI; (b) culturing the cell under conditions in which a POI-cell surface capture molecule complex is formed intracellularly and expressed on the cell surface; © contacting the cell with a detection molecule capable of binding the POI, wherein the surface-displayed POI is detected; and (d) isolating the detected cell.

A method of detecting and isolating a eukaryotic cell that produces a secreted protein of interest (POI), comprising: (a) transfecting a cell with a nucleic acid molecule that encodes a secreted POI comprising an Fc domain; (b) transfecting the cell with a nucleic acid molecule that encodes a cell surface capture molecule capable of binding an Fc domain; © culturing the cell under conditions in which a POI-cell surface capture molecule complex is expressed on the cell surface; (d) contacting the cell with a detection molecule capable of binding the POI, wherein the surface-displayed POI is detected; (e) isolating the detected cell.

View this patent on the USPTO website.