Cell lines for high level production of protein selected for rapid proliferation
AssigneeCHO Plus, Inc.
InventorLawrence Forman
Life of this patent
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Abstract
This disclosure provides improved cell lines for manufacture of protein, considerably reducing the cost of commercial production. The cell lines are obtained by selecting cells from a mixed population for one or more characteristics that support cell growth, compared with other cells in the starting mixture. Particularly effective producer cell lines can be obtained by preparing the cells for functional selection by making cell hybrids. A gene encoding a therapeutic protein of interest may be transfected into the cells before or after one or more cycles of fusion and selection. Depending on the protein product being expressed, cell lines may be obtained that produce eight grams or more of protein per liter of culture fluid.
Claims17
A method of obtaining a derivative cell line for industrial production of a therapeutic protein, the method comprising:
- (a) providing a starting culture of eukaryotic cells that has been adapted to produce a protein encoded in a transgene;
- (b) forming hybrid cells, each comprising two or more cells from the starting culture;
- (c) selecting and recovering hybrid cells that proliferate more rapidly than other hybrid cells formed in step (b) and cells in the starting culture;
- and
- (d) establishing said derivative cell line from the hybrid cells selected and recovered in step (c).
A method of obtaining a hybrid cell line that is adapted for industrial production of protein encoded in a transgene, the method comprising:
- (a) providing a starting mixture of eukaryotic cells;
- (b) forming hybrid cells, each comprising two or more cells from the starting mixture;
- then
- (c) selecting and recovering hybrid cells that proliferate more rapidly than other hybrid cells formed in step (b);
- and
- (d) establishing said hybrid cell line from the hybrid cells selected and recovered in step (c),
- wherein the starting mixture provided in step (a) consists essentially of cells from a single cell line.
A method of obtaining a hybrid cell line for production of a therapeutic protein, the method comprising:
- (a) providing a starting culture of cells or progeny thereof made by fusing cells together to form a population of hybrid cells and enriching the hybrid cells for a higher concentration of endoplasmic reticulum and/or Golgi apparatus compared with other hybrid cells in the population;
- then
- (b) selecting and recovering hybrid cells from the starting culture that proliferate more rapidly from other cells in the culture by successive dilution,
- wherein the successive dilution comprises culturing the hybrid cells in the starting culture at a starting concentration, diluting and reculturing the hybrid cells periodically, and increasing the extent of the diluting and/or decreasing the time of the reculturing so as to dilute out slower growing cells and enrich for faster growing cells.
Dependencies are inferred from the claim text.