Methods and compositions for increasing protein yield from a cell culture
AssigneeClonexDevelopment, Inc.
InventorsTodd Bucciarelli, Victor Levenson, Thomas Primiano
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Abstract
Disclosed herein are compositions and methods for increasing protein production from a cell culture. By switching the cells from a replicative to a productive state (RP switch), protein biosynthesis is extended. The productive state is a pseudo-senescent state. This pseudo-senescent state can be induced by transforming the cells with a vector expressing a cell cycle inhibitor. Expression of the cell cycle inhibitor within the cell, because it does not cause cell death, allows for cells to be maintained in culture for longer periods. The invention allows for controlled enhanced protein biosynthetic productivity of cell lines for commercial and research purposes.
Claims13
A method of increasing yield of a protein from a cell culture, comprising:
- (a) transfecting a cell of the cell culture with at least a first and a second expression vector, wherein the first expression vector encodes a Cip/Kip cyclin-dependent kinase inhibitor family member and the second expression vector encodes an INK4 cyclin-dependent kinase inhibitor family member and wherein each expression vector comprises an inducible transcription regulation element comprising at least two tetracycline operator elements;
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- (b) collecting a protein fraction from the cell culture, wherein the cyclin-dependent kinase inhibitors induce a pseudo-senescent state, characterized by increased protein synthesis, in the cell.
Dependencies are inferred from the claim text.