Integrated cell culture protein purification system for the automated production and purification of proteins

A process for recovering a product secreted by cells growing in a cell culture and purification system, comprising the steps of:

• a. culturing cells grown in a cell culture and purification system, the system consisting essentially of:
• (i) a cell culture subunit for culturing cells;
• (ii) a purification subunit linked to said cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium, said purification subunit comprising a self-contained plug-in cartridge adapted for ready attachment to or removal from said system;
• and
• (iii) means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit, and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• b. extensively washing the cartridge with a wash buffer to remove contaminating protein;
• c. eluting the cartridge with amount elution medium to obtain purified product;
• and
• d. regenerating the cartridge with consecutive washes with wash buffer and subculture medium for a subsequent absorption cycle;
• the process separating said product from other molecules of substantially equivalent molecular weight by specific affinity interactions and removing one immunoglobulin molecule from another immunoglobulin molecule of different binding specificity.

A discontinuous process for recovering and purifying a product secreted by cells grown in a cell culture and purification system, comprising the steps of:

• a. culturing cells grown in a cell culture and purification system, the system consisting essentially of:
• (i) a cell culture subunit for culturing cells;
• (ii) a purification subunit linked to said cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium, said purification subunit comprising a self-contained plug-in cartridge adapted for ready attachment to or removal from said system;
• and
• (iii) means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit, and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• b. withdrawing spent culture medium from said culture subunit by using a pump;
• c. directing said spent medium onto said purification cartridge comprising an absorption medium;
• d. removing unbound cell culture medium constituents by a wash process;
• and
• e. eluting bound products from said purification cartridge to obtain purified product;
• said purification means separating said product from other molecules of substantially equivalent molecular weight by specific affinity interactions and removing one immunogobulin molecule from another immunoglobulin molecule of different binding specificity.

An integrated cell culture and purification system for producing a purified cell culture product, consisting essentially of:

• a. a cell culture subunit for culturing cells;
• b. a purification subunit linked to the cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium;
• and
• c. means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit, and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• said purification subunit separating said product from other molecules of substantial molecular weight by a chromatographic method selected from ion exchange and hydrophobic interaction chromatography and removing one immunoglobulin molecule from another immunoglobulin molecule of different binding specificity.

A process for recovering a product secreted by cells growing in a cell culture and purification system, comprising the steps of:

• a. culturing cells grown in cell culture and protein purification system, the system consisting essentially of:
• (i) a cell culture subunit for culturing cells;
• (ii) a purification subunit linked to said cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium, said purification subunit comprising a self-contained plug-in cartridge adapted for ready attachment to or removal from said system;
• and
• (iii) means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit, and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• b. extensively washing the cartridge with a wash buffer to remove contaminating protein;
• c. eluting the cartridge with a mild elution buffer to obtain purified product;
• and
• d. regenerating the cartridge with consecutive washes with wash buffer and subculture medium for a subsequent absorption cycle;
• the process separating said product from other molecules of substantially equivalent molecular weight by a chromatographic method selected from ion exchange and hydrophobic interaction chromatography and removing one immunoglobulin molecule from another immunoglobulin molecule of different binding specificity.

A discontinuous process for recovering and purifying a product secreted by cells growing in a cell culture and purification system, comprising the steps of:

• a. culturing cells grown on a cell culture and purification system, the system consisting essentially of:
• (i) a cell culture subunit for culturing cells;
• (ii) a purification subunit linked to said cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium, said purification subunit comprising a self-contained plug-in cartridge adapted for ready attachment to or removal from said system;
• and
• (iii) means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit, and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• b. withdrawing spent culture medium from said culture subunit by using a pump;
• c. directing said spent medium onto said purification cartridge comprising an absorption medium;
• d. removing an unbound cell culture medium constituent by a wash process;
• e. eluting bound product from said purification cartridge to obtain purified product;
• said purification means separating said product from other molecules of substantially equivalent molecular weight by a chromatographic method selected from ion exchange and hydrophobic interaction chromatography and removing one immunoglobutin molecule from another immunoglobulin molecule of different binding specificity.

An integrated cell culture and purification system for producing a purified cell culture product, consisting essentially of:

• a. a cell culture subunit for culturing cells;
• b. a purification subunit linked to the cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium, the solid-phase medium being substantially free of leachables toxic to the cells being cultured;
• and
• c. means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• said purification subunit separating said product from other molecules of substantially equivalent molecular weight by specific affinity interactions and removing one immunoglobulin molecule from another immunoglobulin molecule of different binding specificity.

An integrated cell culture and purification subunit for producing a purified cell culture product, consisting essentially of:

• a. a cell culture subunit for culturing cells;
• b. a purification subunit linked to the cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium, the solid-phase medium providing at least one month of continuous cell culture without significant toxic effects on the cells being cultured;
• and
• c. means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• said purification subunit separating said product from other molecules of substantially equivalent molecular weight by specific affinity interactions and removing one immunoglobulin molecule from another immunoglobulin molecule of different binding specificity.

An integrated cell culture and purification system for producing a purified cell culture product, consisting essentially of:

• a. a cell culture subunit for culturing cells;
• b. a purification subunit linked to the cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium, the solid-phase medium being substantially free of leachables toxic to the cells being cultured;
• and
• c. means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• said purification subunit separating said product from other molecules of substantially equivalent molecular weight by a chromatographic method selected from ion exchange and hydrophobic interaction chromatography and removing one immunoglobulin molecule from another immunoglobulin molecule of different binding specificity.

An integrated cell culture and purification system for producing a purified cell culture product, consisting essentially of:

• a. a cell culture subunit for culturing cells;
• b. a purification subunit linked to the cell culture subunit adapted to remove said product from a culture fluid in which the cells are cultured by purification means comprising a solid-phase medium, the solid-phase medium providing at least one month of continuous cell culture without significant toxic effects on the cells being cultured;
• and
• c. means for circulating said culture fluid from said cell culture subunit to said purification subunit;
• said cell culture subunit, said purification subunit, and said means for circulating being integrated in a single unit and contained in a sterile environment to purify said product in a sterile environment without removing said culture fluid from said integrated cell culture and purification system;
• said purification subunit separating said product from other molecules of substantially equivalent molecular weight by a chromatographic method selected from ion exchange and hydrophobic interaction chromatography and removing one immunoglobulin molecule from another immunoglobulin molecule of different binding specificity.