Cell culture method to increase cell biomass, productivity and protein titer

A cell culture process for production of a recombinant protein, comprising:

• a) culturing cells in a primary fermenter and/or bioreactor which produces a desired amount of biomass in cell culture;
• b) feeding the cells in the primary fermenter with feeding medium containing glucose;
• c) transferring the cultured cells from the primary fermenter into a secondary fermenter which is connected to each other;
• and
• d) feeding the cells in the secondary fermenter with feeding medium;
• e) expressing the recombinant protein in the secondary fermenter;
• wherein transferring of the cultured cells into the secondary fermenter is performed when biomass level is about 150 g/L to about 230 g/L.

A cell culture process for production of protein comprising:

• a) culturing cells in a primary fermenter;
• b) feeding glucose in the primary fermenter under suitable conditions;
• c) transferring the cells from the primary fermenter to a secondary fermenter to obtain desired yield of protein of interest;
• and
• d) expressing the protein of interest in the secondary fermenter;
• wherein cell density in the primary fermenter and protein titer in the secondary fermenter is increased;
• wherein the induction is performed in the secondary fermenter to express and harvest the protein of interest from the secondary fermenter.

A cell culture process for production of recombinant protein comprising:

• a) culturing cells in a serum free, chemically defined complex media, wherein culturing of cells from a seed culture to a first liquid cell culture;
• b) expanding the cells that produce recombinant protein, wherein expanding of the cell culture from the first liquid cell culture to a second liquid cell culture;
• and
• c) harvesting the recombinant protein from the second liquid cell culture;
• wherein the first liquid cell culture is transferred to the second liquid cell culture after achieving biomass about 150 g/l to about 230 g/l.

A cell culture process for production of protein comprising:

• a) culturing cells in a primary fermenter;
• b) feeding glucose in the primary fermenter under suitable conditions;
• c) transferring the cells from the primary fermenter to a secondary fermenter to obtain desired yield of protein of interest;
• and
• d) expressing the protein of interest in the secondary fermenter;
• wherein feeding of glucose is performed at suitable time during the culture in the primary fermenter to achieve cell density at least OD600 120 to OD600 140;
• wherein the induction is performed in secondary fermenter to express and harvest the protein of interest from the secondary fermenter.

A method for increasing productivity of a cell culture process comprising:

• a) culturing cells in a primary fermenter which produces a desired amount of biomass in cell culture under conditions that allow for protein production;
• b) feeding the cells in the primary fermenter with feeding medium containing glucose;
• c) transferring the cultured cells from the primary fermenter into a secondary fermenter which is connected to each other;
• and
• d) feeding the cells in the secondary fermenter with feeding medium;
• e) expressing the recombinant protein in the secondary fermenter;
• wherein transferring of the cultured cells into the secondary fermenter is performed when biomass level is about 150 g/L to about 230 g/L.

A method for increasing titer of a recombinant protein in a cell culture process comprising:

• a) culturing cells in a primary fermenter which produces a desired amount of biomass in cell culture under conditions that allow for protein production;
• b) feeding the cells in the primary fermenter with feeding medium containing glucose;
• c) transferring the cultured cells from the primary fermenter into a secondary fermenter which is connected to each other;
• and
• d) feeding the cells in the secondary fermenter with feeding medium;
• e) expressing the recombinant protein in the secondary fermenter;
• wherein transferring of the cultured cells into the secondary fermenter is performed when biomass level is about 150 g/L to about 230 g/L.

A process for cell culture to produce a recombinant protein, cell culture process comprising:

• a) culturing cells in a primary fermenter which produces a desired amount of biomass in cell culture under conditions that allow for protein production;
• b) feeding the cells in the primary fermenter with feeding medium containing glucose;
• c) transferring the cultured cells from the primary fermenter into a secondary fermenter which is connected to each other;
• and
• d) feeding the cells in the secondary fermenter with feeding medium;
• e) expressing the recombinant protein in the secondary fermenter;
• wherein transferring of the cultured cells into the secondary fermenter is performed when biomass level is about 150 g/L to about 230 g/L;
• wherein the cell density, cell biomass, titer and total inclusion bodies are increased.