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A corn active peptide additive for cell culture medium

A technology of active peptides and additives, applied in the field of cell culture medium additives, can solve the problems of cell apoptosis, destroy cell membrane structure, change cell shape, etc., and achieve the effects of low endotoxin content, promotion of cell proliferation, and high enzymatic hydrolysis efficiency.

Active Publication Date: 2018-05-04
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

[0004] Corn active peptides are rich in nutrients, and are easily contaminated by bacterial microorganisms during their processing and storage, thereby producing a large amount of bacterial endotoxins. When used as cell culture additives, they will change cell morphology, destroy cell membrane structure, induce cell apoptosis, etc. Adversely affect the growth of cells cultured in vitro
In addition, because different processing techniques have a great impact on the composition of the prepared corn active peptide products, and the composition of animal serum is relatively complex, many components are still unclear, so it is impossible to determine the existence or existence of certain components or certain components. Whether the deletion will have adverse effects on the growth of cells, which will cause some difficulties in the application of corn active peptides in serum-free medium supplements

Method used

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  • A corn active peptide additive for cell culture medium
  • A corn active peptide additive for cell culture medium
  • A corn active peptide additive for cell culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1. Enzymatic treatment

[0050] Add 12 times deionized water to corn yellow powder with protein content ≥ 60%, mix well, heat up to 90°C, keep warm for 15-20 minutes, and when the temperature is lowered to 55°C, adjust the pH value of the protein solution to 7-8 , adding a compound non-specific protease mixed with neutral protease NEUTRASE O.D.L (Novozymes) and alkaline protease ALKALINE PROTEASECONCENTRATE (DSM Company) to the protein solution with an enzyme amount of 4000 units per gram of protein (wherein The enzyme amount of NEUTRASE O.D.L and alkaline protease ALKALINE PROTEASECONCENTRATE is 2000 units), and the first enzymolysis treatment was incubated at 55°C for 2.5 hours to obtain the first enzymolysis solution; the pH value of the first enzymolysis solution was adjusted to After 7.5 to 8.5, add carboxypeptidase ACCELERZYMECPG (DSM company) to the first enzymatic hydrolysis solution with an enzyme amount of 2000 units per gram of protein, and incubate at 50°C f...

Embodiment 2

[0069] The enzymatic hydrolysis treatment is specifically as follows: add 9 times deionized water to corn yellow powder with protein content ≥ 60%, mix well, heat up to 90-95°C, keep warm for 55-60 minutes, and then cool down to 50-55°C , adjust the pH of the protein solution to be 7-8, add papain (Nanning Pangbo Bioengineering Co., Ltd.) and bromelain (Nanning Pangbo Biological Engineering Co., Ltd.) Company) mixed compound non-specific protease (the enzyme amounts of papain and bromelain are 1000 units and 1500 units respectively), and the first enzymolysis treatment was incubated at 50-55°C for 1.5 hours to obtain the first enzymolysis solution; after adjusting the pH value of the first enzymolysis solution to 8-9, then add carboxypeptidase Validase FP Conc. (DSM company) and Flavor protease FLAVOURZYME500MG (Novozymes) is a composite specific protease (in which the enzyme amount of carboxypeptidase ACCELERZYME CPG and flavor protease FLAVOURZYME500MG are both 1500 units), ...

Embodiment 3

[0072] The enzymatic hydrolysis treatment is specifically: add 10 times deionized water to corn yellow powder with protein content ≥ 60%, mix well, heat up to 95°C, keep warm for 35-40 minutes, cool down to 45-50°C, adjust The pH value of the protein solution is 7-8, and the neutral protease Validase BNP-L (DSM company), alkaline protease Alcalase2.4L (Novozymes) is added to the protein solution with an enzyme amount of 6000 units per gram of protein. company), papain (Nanning Pangbo Bioengineering Co., Ltd.) and bromelain (Nanning Pangbo Biological Engineering Co., Ltd.) , papain and bromelain (the enzyme amounts of 2000 units, 2000 units, 1000 units and 1000 units respectively) were incubated at 45-50°C for the first enzymolysis treatment for 3 hours to obtain the first enzymolysis solution; After the pH value of the enzymolysis solution is 5.5, add flavor protease MAXZPRO XF (DSM company) to the first enzymolysis solution with an enzyme amount of 2000 units per gram of prot...

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Abstract

The invention provides a corn active peptide additive for cell culture medium, wherein the oligopeptides with a molecular weight less than 1000 Daltons in the corn active peptide additive account for ≥90% of the total protein, and the oligopeptides are at least Including one or more of AP, SAP, PAL, VNAP, PSSQ, TQPGPQ. The corn active peptide additive of the present invention can be compounded with various basal media for serum-free culture of various 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 corn active peptide additive for a cell culture medium. Background technique [0002] Culture medium is the most direct and important environmental factor that determines the growth and metabolism of cultured cells in vitro. Animal cell culture media usually contain animal-derived additives such as serum, and with the outbreak of mad cow disease worldwide, the use of serum and other animal-derived additives has brought infectious spongiform encephalopathy (TSEs) viruses and other dangerous substances to the biopharmaceutical industry The security risk has risen sharply. Drug regulators around the world are asking drug manufacturers to reduce the use of bovine-derived ingredients in pharmaceutical production, and relevant legislatures are increasingly opposed to the use of animal-derived ingredients such as bovine serum in pharmaceuticals. The US Food and Drug Administration (...

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
CPCC12P21/06C07K5/06026C07K5/081C07K5/0819C07K5/0823C07K5/1008C07K5/101C07K5/1024C07K7/06
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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