Method of identifying neonatal calf serum quality

A new-born bovine serum and quality technology, applied in the field of identification, can solve the problems of low accuracy of identification results, complicated operation of identification methods, poor repeatability, etc., and achieve the effects of easy popularization and application, simple method and prolonged lifespan.

Active Publication Date: 2016-05-11
华北制药金坦生物技术股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to provide a method for identifying the quality of newborn bovine serum to solve the problems of cumbersome operation, long time consumption, low accuracy and poor repeatability of identification results in existing identification methods

Method used

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  • Method of identifying neonatal calf serum quality
  • Method of identifying neonatal calf serum quality
  • Method of identifying neonatal calf serum quality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Samples: Newborn bovine serum, fetal bovine serum, and calf serum, which are identified as good performance and quality in the cell culture stage for regular annual use by manufacturer A.

[0037] Reagents used:

[0038] Reagents used for the detection of total protein content: NaOH, NaCl, Na2CO3, CuSO4, potassium sodium tartrate (Sinopharm Chemical Reagent Co., Ltd. AR), phenol reagent (SIGMAAR), standard substance: serum albumin (bovine) (China Institute for Food and Drug Control);

[0039] Reagents for electrophoresis detection: 30% acrylamide / methylenebisacrylamide 37.5:1 (Yisheng Bio), TEMED (GE), SDS (SIGMA), Coomassie brilliant blue staining solution (SIGMA);

[0040] Reagents used for lipid content detection: ethanol, sulfuric acid, phosphoric acid, vanillin (Sinopharm Chemical Reagent Company AR).

[0041] Instruments used:

[0042] UV spectrophotometer: PerkinElmerLambda25; electrophoresis tank: BIO-RADMINI2; optical density scanner: BIO-RADGS-800CalibratedDen...

Embodiment 2

[0062] Bovine serum samples: 16 batches of bovine serum from 7 different domestic manufacturers and 2 batches of bovine serum from 1 foreign manufacturer were selected, totaling 18 batches of serum from 8 manufacturers, and numbered from 1 to 18.

[0063] The reagents and instruments used were the same as those in Example 1.

[0064] Each sample was tested as follows:

[0065] Detection of total protein content: the same as in Example 1.

[0066] Detection of albumin and impurity protein content:

[0067] Gel preparation: the gel concentration of separating gel is 15%, and the gel concentration of stacking gel is 4.5%.

[0068] Add sample: use BIO-RADMINI2 electrophoresis tank and 10-well sample comb; after the stacking gel is polymerized, remove the sample comb, add bovine serum sample, and add 20 μL of sample to each well.

[0069] Electrophoresis and staining: Turn on the power and start electrophoresis; after electrophoresis, use Coomassie brilliant blue for overnight s...

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Abstract

The invention provides a method of identifying neonatal calf serum quality, comprising the following steps: full inspecting qualification of a sample to be identified, dependent on the methods in Chinese Pharmacopeia III appendix; detecting total protein content of the sample to be identified, dependent on 'Lowry process' in 'protein determination' from Chinese Pharmacopeia III appendix; subjecting the sample to be identified to non-reducing electrophoretic detection dependent on 'SDS-polyacrylamide gel electrophoresis' in 'electrophoresis' from Chinese Pharmacopeia III appendix, quantitatively scanning and analyzing electrophoresis results, and calculating albumin content of the sample to be identified; measuring lipid content of the sample to be identified by using vanillin development process, wherein the measured total protein content is 4.0-4.5 g / 100ml, the result for quantitative scanning of albumin is 1.0-1., and the sample to be identified, having a lipid content of 5.5-6.5 mg / ml is qualified neonatal calf serum. The invention enables quick and effective screening of high-quality neonatal calf serum and solves the problem that an existing screening method is time consuming and labor consuming.

Description

technical field [0001] The invention relates to an identification method, in particular to a method for identifying the quality of newborn bovine serum. Background technique [0002] The development of modern biotechnology is inseparable from cell culture, and the quality of the medium is the key to cell culture. As the most used natural medium in cell culture, bovine serum is rich in nutrients necessary for cell growth. It not only provides cells with the factors needed for adherence and spreading, but also buffers pH, protects cells from damage, and detoxifies. Therefore, bovine serum is one of the important raw materials in pharmaceutical biotechnology products. In the process of antigen production, newborn bovine serum is often added for the growth and expression of CHO cells. If the serum quality is not good, it will not only affect the growth and expression of cells, but also affect the subsequent purification yield and the quality of the antigen stock solution. The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447G01N21/33
CPCG01N21/33G01N27/447
Inventor 张卫婷王炜荀英李潜张亚丽洪庆福谭丽霞李岩异张红霞刘海侠戴碧璇臧文伟高珊
Owner 华北制药金坦生物技术股份有限公司
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