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US 12,590,880 B2Granted
Flare (flow cytometry attenuated reporter expression) technology for rapid bulk sorting
AssigneeGenzyme Corporation
InventorsVictor R. Cairns, Jose Ignacio Sancho Chavida, Christine DeMaria
Life of this patent
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Abstract
Provided herein are methods and compositions for batch production of producer cells using fluorescence activated cell sorting (FACS). In some aspects, the disclosure provides a drug-selection-free method for batch production of producer cells using FACS. Such batch production methods and compositions can be further utilized to generate clonal populations of producer cells, e.g., for large-scale manufacturing of a polypeptide of interest.
Claims23
A method of increasing viability of cells after fluorescence activated cell sorting (FACS), the method comprising:
- using a first round of FACS to select based on a viability marker a first sub-population of cells from a population of cells expressing a target polypeptide, and
- using a second round of FACS to select based on the viability marker a second sub-population of cells from the first sub-population of cells,
- wherein the first round and second round of FACS occur within 8 hours of each other, and wherein the viability marker is a forward scatter/side scatter population differentiation and/or a propidium iodide stain.
Dependencies are inferred from the claim text.