EARLY POST-TRANSFECTION ISOLATION OF CELLS (EPIC) FOR BIOLOGICS PRODUCTION

摘要

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.

主权项

1. 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; (b) isolating from the transfected host cells, within 2 to 15 days of transfection, a sub-population of early-expressing transfected host cells which express the selectable polypeptide; and (c) expanding the sub-population of transfected host cells, thereby producing a population of producer cells expressing the target polypeptide.