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Cell culture medium and method for producing protein

A technology of cell culture and culture medium, applied in the field of biopharmaceuticals, which can solve the problems of potential risks in the stability of product quality between batches, complex components of hydrolyzate, and difficulty in controlling batch-to-batch consistency, and achieve good product consistency and easy quality. Effects of management and productivity improvement

Inactive Publication Date: 2018-03-06
SICHUAN KELUN BIOTECH BIOPHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the composition of the hydrolyzate is complex, and the batch-to-batch consistency is difficult to control, so there is a potential risk to the batch-to-batch stability of product quality

Method used

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  • Cell culture medium and method for producing protein
  • Cell culture medium and method for producing protein
  • Cell culture medium and method for producing protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1, cell culture medium

[0060] 1. Preparation of basal medium

[0061] The formulation of the basal medium: CD FortiCHO AGT (Thermo Fisher Company), 12.5g / L; Ex-Cell302 (Sigma-Aldrich Company), 10.8g / L.

[0062] Preparation method: Take 80% of the preparation volume of water for injection in the liquid preparation container, weigh the Ex-cell302 dry powder and CD FortiCHO AGT dry powder required for the culture medium, add them to the liquid preparation container, and stir for 30 minutes. Adjust the pH to 6.9-7.0 with 5M sodium hydroxide, and continue stirring for about 10 minutes. Dilute to the prepared volume, filter and aliquot. Verify sterility after filtration and check filter integrity. Store in a refrigerator at 4°C.

[0063] 2. Preparation of feed medium

[0064] The formula and preparation method of the feed medium are as follows:

[0065] Table 1: Recipe of feed medium

[0066]

[0067]

[0068] Preparation method: Take 80% of the prep...

Embodiment 2

[0070] A cell culture medium comprising basal medium and feed medium,

[0071] The formulation of the basal medium: CD FortiCHO AGT, 5g / L; Ex-Cell 302, 5g / L.

[0072] Formula of feed medium: CD EfficientFeed C AGT, 40g / L; Cell Boost 5, 20g / L; Glutamic acid 0.1g / L; Tyrosine 0.5g / L; Cysteine ​​0.2g / L; Amino acid 0.1g / L; PF68 1g / L.

Embodiment 3

[0074] A cell culture medium comprising basal medium and feed medium,

[0075] The formulation of the basal medium: CD FortiCHO AGT, 20g / L; Ex-Cell 302, 15g / L.

[0076] Formula of feed medium: CD EfficientFeed C AGT, 80g / L; Cell Boost 5, 70g / L; Glutamic acid 0.8g / L; Tyrosine 1.2g / L; Cysteine ​​1g / L; Tryptophan Acid 0.8g / L; threonine 0.3g / L; phenylalanine 0.4g / L; PF68 2g / L.

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PUM

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Abstract

The invention belongs to the field of biological pharmacy, and provides a cell culture medium and a method for producing protein by using the culture medium to culture cells. According to the cell culture medium, amino acid optimization is carried out on the basis of a culture medium limited by chemical components, so that the protein expression capacity of a cell is improved. The method providedby the invention is simple in process control and is beneficial to large-scale production.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals and relates to a method for producing protein through cell culture. Background technique [0002] Cetuximab (cetuximab, trade name Erbitux) is an anti-EGFR human-mouse chimeric monoclonal antibody developed by ImClone Systems, which is used to treat metastatic colorectal cancer and head and neck cancer with high expression of EGFR. cancer. The host cell expressing cetuximab is murine myeloma cell SP2 / 0. In addition to the modification of non-human NGNA (N-glycolylneuraminic acid, N-glycolylneuraminic acid), the protein expressed by SP2 / 0 cells also has a small amount of mouse α-(1,3) galactose residues, which can Triggers a severe immune response in the body. Cetuximab has clinically caused severe adverse events such as anaphylactic shock due to its murine-derived "α-(1,3) galactose residue" structure, which limits its use to a certain extent. [0003] Currently, more than 70% of antibody pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C07K16/28
CPCC07K16/2863C12N5/0682C12N2500/32C12N2510/02
Inventor 薛彤彤杨秋艳李明雄王帅坤谢冰松刘立平王利春王晶翼
Owner SICHUAN KELUN BIOTECH BIOPHARMACEUTICAL CO LTD
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