Isolating cells expressing secreted proteins
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsJames Fandl, Neil Stahl, Gang Chen, George Yancopoulos
Life of this patent
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Abstract
A method of detecting and isolating cells that produce any secreted protein of interest (POI), comprising: a) constructing a cell line transiently or stably expressing a cell surface capture molecule, which binds the POI, by transfecting the cell line with a nucleic acid that encodes such cell surface capture molecule; b) transfecting said cell simultaneously or subsequently with a second nucleic acid that encodes a POI wherein such POI is secreted; c) detecting the surface-displayed POI by contacting the cells with a detection molecule, which binds the POI; and d) isolating cells based on the detection molecule.
Claims18
A method of detecting and isolating cells that produce a secreted protein of interest (POI), comprising:
- a) constructing a cell line transiently or stably expressing a cell surface capture molecule, which binds the POI, by transfecting the cell line with a nucleic acid that encodes such cell surface capture molecule;
- b) transfecting said cell simultaneously or subsequently with a second nucleic acid that encodes a POI wherein such POI is secreted;
- c) detecting the surface-displayed POI by contacting the cells with a detection molecule, which binds the POI;
- d) isolating cells based on the detection molecule.
A method of detecting and isolating cells that produce any secreted protein of interest (POI), comprising:
- a) constructing a cell line transiently or stably expressing a cell surface capture molecule, which binds the POI, by transfecting the cell line with a nucleic acid that encodes such cell surface capture molecule;
- b) detecting a cell from (a) that expresses said cell surface capture molecule;
- c) isolating and culturing the cell detected in (b);
- d) transfecting said cell in (c) simultaneously or subsequently with a second nucleic acid that encodes a POI wherein such POI is secreted;
- e) detecting the surface-displayed POI by contacting the cells with a detection molecule, which binds the POI;
- f) isolating cells based on the detection molecule.
A method of detecting and isolating cells that produce a POI, comprising:
- a) detecting a cell that expresses said cell surface capture molecule in high yield;
- b) isolating and culturing the cell detected in (a);
- c) transfecting said cell in (b) with a nucleic acid that encodes a POI wherein such POI is secreted;
- d) detecting the surface-displayed POI by contacting the cells with a detection molecule which binds the POI;
- e) isolating cells based on the detection molecule.
Dependencies are inferred from the claim text.