Anti-vegf protein compositions and methods for producing the same
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsShunhai Wang, Ning Li, Hunter Chen, Amardeep Singh Bhupender Bhalla, Shawn M. Lawrence, Amy Johnson, Meghan Casey, Jaimie Grapel
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Abstract
The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions.
Claims30
A method of preparing a prefilled syringe comprising aflibercept at a concentration of about 8 mg/0.07 mL, wherein said aflibercept is produced in a cell culture media including a chemically defined media (CDM) by:
- (a) providing a host cell genetically engineered to express aflibercept;
- (b) culturing said host cell in said cell culture media including said CDM under conditions suitable in which said host cell expresses said aflibercept wherein the cumulative concentration of nickel in said CDM is less than or equal to 0.4 μM or about 0.4 μM and one or more of the following:
- i. the cumulative concentration of iron in said CDM is less than or equal to 55.0 μM;
- ii. the cumulative concentration of copper in said CDM is less than or equal to 0.8 μM;
- iii. the cumulative concentration of zinc in said CDM is less than or equal to 56.0 μM;
- iv. the cumulative concentration of cysteine in said CDM is less than or equal to 10.0 mM, and v. said CDM includes anti-oxidants where the cumulative concentration of an antioxidant is about 0.001 mM to about 10.0 mM for any single anti-oxidant;
- and (c) harvesting aflibercept produced by said host cell.
A method of preparing a prefilled syringe comprising aflibercept at a concentration of about 8 mg/0.07 mL, wherein said aflibercept is produced in a cell culture media including a chemically defined media (CDM) by:
- (a) providing a host cell genetically engineered to express aflibercept;
- (b) culturing said host cell in said CDM under conditions suitable in which said host cell expresses said aflibercept wherein the cumulative concentration of nickel in said CDM is about 0.4 μM and one or more of the following:
- i. the cumulative concentration of iron in said CDM is less than or equal to 55.0 μM;
- ii. the cumulative concentration of copper in said CDM is less than or equal to 0.8 μM;
- iii. the cumulative concentration of zinc in said CDM is less than or equal to 56.0 μM;
- iv. the cumulative concentration of cysteine in said CDM is less than or equal to 10.0 mM;
- and v. said CDM includes anti-oxidants where the cumulative concentration of an antioxidant is about 0.001 mM to about 10.0 mM for any single anti-oxidant;
- and (c) harvesting aflibercept produced by said host cell, wherein said aflibercept produced by said host cell comprises one or more aflibercept variants, wherein said variants have at least one oxidized amino acid residue selected from histidine, tryptophan, and a combination thereof.
A method of preparing a prefilled syringe comprising aflibercept at a concentration of about 110 mg/mL to about 120 mg/mL, wherein said aflibercept is produced in a cell culture media including a chemically defined media (CDM) by:
- (a) culturing a host cell in said CDM under conditions suitable in which said host cell expresses said aflibercept wherein the cumulative concentration of nickel in said CDM is less than or equal to 0.4 μM or about 0.4 μM and one or more of the following:
- i. the cumulative concentration of iron in said CDM is less than or equal to 55.0 μM;
- ii. the cumulative concentration of copper in said CDM is less than or equal to 0.8 μM;
- iii. the cumulative concentration of zinc in said CDM is less than or equal to 56.0 μM;
- iv. the cumulative concentration of cysteine in said CDM is less than or equal to 10.0 mM;
- and v. said CDM includes anti-oxidants where the cumulative concentration of an antioxidant is about 0.001 mM to about 10.0 mM for any single anti-oxidant;
- and (b) harvesting aflibercept produced by said host cell.
Dependencies are inferred from the claim text.