Early post-transfection isolation of cells (EPIC) for biologics production
AssigneeGenzyme Corporation
InventorsVictor R. Cairns, Christine DeMaria, Jason Vitko
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Abstract
Provided herein are methods for selecting a population of cells expressing a target polypeptide. In some aspects, the disclosure provides methods for sorting and selecting populations of transfected host cells based on their early expression of a selectable polypeptide. In certain embodiments, the sorting is performed using fluorescence-activated cell sorting or magnetic-activated cell sorting based on the selectable polypeptide. Such selection methods can be further utilized to generate clonal populations of producer cells, e.g. for large-scale manufacturing of a target polypeptide of interest.
Claims15
A method of producing a population of producer cells expressing a target polypeptide, the method comprising:
- (a) transfecting host cells with one or more vectors that encode one or more mRNAs, wherein the one or more mRNAs encode a selectable polypeptide and the target polypeptide, wherein the mRNA encoding the target polypeptide and the mRNA encoding the selectable polypeptide are both encoded on one vector;
- (b) isolating from the transfected host cells, within 2 to 5 days of transfection, a sub-population of early-expressing transfected host cells which express the selectable polypeptide, wherein the isolating employs magnetic activated cell sorting (MACS) or fluorescence activated cell sorting (FACS) on the selectable polypeptide;
- and
- (c) expanding the isolated sub-population of early-expressing transfected host cells, thereby producing a population of producer cells expressing the target polypeptide,
- wherein the population of producer cells expresses an enhanced level of the target polypeptide compared to an unselected population of the transfected host cells, and
- wherein steps (b) and (c) are each performed in drug-selection-free medium.
Dependencies are inferred from the claim text.