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A kind of wheat active peptide additive for cell culture medium

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

Active Publication Date: 2017-12-01
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 kind of wheat active peptide additive for cell culture medium
  • A kind of wheat active peptide additive for cell culture medium
  • A kind of wheat active peptide additive for cell culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] 1. Enzymatic treatment

[0051] Add 12 times deionized water to wheat gluten with protein content ≥ 80%, mix well, heat up to 90°C, keep warm for 15-20 minutes, and adjust the pH value of the protein solution when the temperature is lowered to 45-50°C To 6.5~7.5, with the enzyme amount of 3000 units per gram of protein, add the compound non-specific protease mixed by neutral protease PROTAMEX (Novozymes Company) and papain (Nanning Pangbo Bioengineering Co., Ltd.) Heterotropic protease (wherein the enzyme amounts of neutral protease PROTAMEX and papain are 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 solution is 7.5 to 8.5, add carboxypeptidase Validase FP Conc. (DSM company) to the first enzymolysis solution with an enzyme amount of 2000 units per gram of protein, and incubate at 45 to 50°C After the second enzymatic h...

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 During ℃, the pH value of regulating protein solution is 7~8, adds in described protein solution with the enzyme amount of every gram protein 3500 units and is made of alkaline protease Novozym37071 (Novozym) and bromelain (Nanning Pangbo Biological Engineering Co., Ltd. Company) mixed compound non-specific protease (the enzyme amounts of alkaline protease Novozym 37071 and bromelain were 2000 units and 1500 units respectively), and the first enzymolysis treatment was incubated at 50-55°C for 2 hours to obtain the second One enzymolysis solution; after adjusting the pH value of the first enzymolysis solution to 8-9, add carboxypeptidase ACCELERZYME CPG (DSM) and Flavor protease FLAVOURZYME 500MG (Novozymes) is a composite specific protease (in which the ...

Embodiment 3

[0073] The enzymatic hydrolysis treatment is specifically as follows: 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 protein solution is 6~7, and in described protein solution, add papain (Nanning Pangbo Biological Engineering Co., Ltd.), Bromelain (Nanning Pangbo Biological Engineering Co., Ltd.) ) and neutral protease ACCELERZYME NP50000 (DSM company) mixed compound non-specific protease (wherein the enzyme amounts of papain, bromelain and neutral protease are respectively 1000 units, 1500 units and 2000 units), incubated at 50°C 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 MAXZPRO XF (DSM company), heat up ...

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Abstract

The invention provides a wheat active peptide additive for cell culture medium, in which the oligopeptides with a molecular weight less than 1000 Daltons account for more than 90% of the total protein by mass, and the oligopeptides are at least Including one or more of VN, TFN, QVSQ, HGAQ, GGDNP, GTCGAG. The wheat active peptide additive of the present invention can be compounded with a variety of basal media for serum-free culture of a variety of animal cells, which not only greatly reduces the cost of cell culture media, but also reduces the cost of animal-derived components. It can also promote cell proliferation, improve cell viability and enhance the 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, and adhesion factors. 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. [00...

Claims

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Application Information

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Patent Type & Authority Patents(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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