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A wheat bioactive peptide additive used for a cell culture medium

A wheat active peptide and additive technology, which is applied to animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of difficult application of wheat active peptides, complex animal serum components, adverse effects on cells, etc., and achieves easy preparation and operation. Simple, low-cost effect

Active Publication Date: 2015-05-06
CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, because different processing techniques have a great influence on the composition of the prepared wheat active peptide products, and the components of animal serum are relatively complex, and many components are still unclear, so it is impossible to determine whether the presence or absence of certain components will cause It has adverse effects on the growth of cells, thus causing certain difficulties in the application of wheat active peptides in serum-free medium additives

Method used

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  • A wheat bioactive peptide additive used for a cell culture medium
  • A wheat bioactive peptide additive used for a cell culture medium
  • A wheat bioactive peptide additive used for a cell culture medium

Examples

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Comparison scheme
Effect test

Embodiment 1

[0050] 1. Enzymatic treatment

[0051] Add 12 times of deionized water to wheat gluten with protein content ≥ 80%, mix evenly, heat up to 90°C, keep it warm for 15-20 minutes, then cool down to 45-50°C, adjust the pH value of the protein solution To 6.5 ~ 7.5, with the enzyme amount of 3000 units per gram of protein, add a compound non-specific protease mixed by neutral protease PROTAMEX (Novozymes) and papain (Nanning Pangbo Bioengineering Co., Ltd.) Heterotropic protease (in which the enzyme amount of neutral protease PROTAMEX and papain is 2000 units and 1000 units respectively), incubate the first enzymolysis treatment at 45-50°C for 3 hours to obtain the first enzymolysis solution; adjust the first enzyme After the pH value of the hydrolyzate is 7.5-8.5, add carboxypeptidase Validase FP Conc. (DSM company) to the first enzymatic hydrolyzate at an enzyme amount of 2000 units per gram of protein, and incubate at 45-50°C After the second enzymatic hydrolysis treatment for 2...

Embodiment 2

[0070] The enzymatic hydrolysis treatment is specifically as follows: add 9 times deionized water to wheat gluten with protein content ≥ 80%, mix well, heat up to 90-95°C, keep warm for 55-60 minutes, then cool down to 50-55°C At ℃, adjust the pH value of the protein solution to 7-8, add alkaline protease Novozym37071 (Novozym) and bromelain (Nanning Pangbo Bioengineering Co., Ltd. Company) mixed compound non-specific protease (among them, the enzyme amounts of alkaline protease Novozym37071 and bromelain are 2000 units and 1500 units respectively), and the first enzymolysis treatment was incubated at 50-55°C for 2 hours to obtain the first Enzymolysis solution: After adjusting the pH value of the first enzymolysis solution to 8-9, add carboxypeptidase ACCELERZYME CPG (DSM) and flavor Protease FLAVOURZYME500MG (Novozymes) is a compound specific protease (in which the enzyme amount of carboxypeptidase ACCELERZYME CPG and flavor protease FLAVOURZYME500MG are both 1000 units), in...

Embodiment 3

[0073] The enzymatic hydrolysis treatment is specifically: add 10 times deionized water to wheat gluten with protein content ≥ 80%, mix well, heat up to 95°C, keep warm for 35-40 minutes, and when the temperature is lowered to 50°C, adjust The pH value of the protein solution is 6-7, and adding papain (Nanning Pangbo Bioengineering Co., Ltd.), bromelain (Nanning Pangbo Biological Engineering Co., Ltd.) to the protein solution with an enzyme amount of 4500 units per gram of protein ) and neutral protease ACCELERZYME NP50000 (DSM company) mixed non-specific protease (in which the enzyme amount of papain, bromelain and neutral protease are 1000 units, 1500 units and 2000 units respectively), and incubated at 50 ℃ The first enzymolysis was treated for 3.5 hours to obtain the first enzymolysis solution; after adjusting the pH value of the first enzymolysis solution to 6, add flavor protease to the first enzymolysis solution with an enzyme amount of 2000 units per gram of protein MA...

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Abstract

A wheat bioactive peptide additive used for a cell culture medium is provided. The mass of oligopeptides having molecular weights lower than 1000 Dalton is not less than 90% of the total protein mass in the additive, and the oligopeptides at least comprise one or more of VN, TFN, QVSQ, HGAQ, GGDNP and GTCGAG. The additive can be blended with various base culture media and used for serum-free culture of a plurality of animal cells. The additive largely reduces the cost of the cell culture medium, reduces pollutions and other problems caused by animal-source components, promotes cell proliferation, improves cell viability and enhances expression of cell products.

Description

technical field [0001] The invention relates to a cell culture medium additive, in particular to a wheat active peptide additive for a cell culture medium. Background technique [0002] Animal cell in vitro culture technology is one of the key upstream technologies of biopharmaceuticals, which can be used in the production of various vaccines, monoclonal antibodies, tumor drugs and other biological products. Cell culture medium is the most direct and important environmental factor that determines the growth and metabolism of cells cultured in vitro. Animal cells are usually cultured with basal medium plus 5-20% serum or serum extracts. The main function is to provide various nutrients, hormones (such as insulin), growth factors, transport proteins, adhesion factors, etc. However, the use of serum and serum extracts has certain limitations due to problems such as high safety risks, unstable batches, and increased purification costs downstream of the production process. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12P21/06C07K1/36C07K1/34C07K1/18
Inventor 蔡木易林峰刘艳张海欣谷瑞增鲁军魏颖
Owner CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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