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Method for detecting recombinant human hepatocyte growth factor activity

A hepatocyte growth factor and detection method technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of time-consuming, complicated operation, and large variation of results, and achieve no radioactive pollution, simple operation, good repeatability

Inactive Publication Date: 2004-08-18
SHENZHEN NEPTUNUS INTERLONG BIO TECHN HLDG
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Problems solved by technology

The above methods mainly have the following deficiencies: (1) The in vivo detection method is complicated to operate, the result varies greatly, and takes a long time, so it is not suitable as a routine method for drug activity detection; (2) The in vitro cell isotope detection method has radiation pollution, large variation, etc. also more complicated

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  • Method for detecting recombinant human hepatocyte growth factor activity
  • Method for detecting recombinant human hepatocyte growth factor activity

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Embodiment Construction

[0029] The NIH3T3 fiber cell line was recovered and frozen, and inoculated into a 10ml culture flask for culture and proliferation. Digest and proliferate NIH3T3 fibroblasts, adjust the cell concentration to 0.5×10 with 1640 culture medium of 5% calf serum 5 cells / ml, spread 96-well plate, 100 μl per well. Set at 37°C, 5% CO 2 Cultivate in an incubator for 24 hours. If the cells are in good growth state, discard the culture medium and add the test substance in groups as follows:

[0030] (1) NIH3T3 fibroblast control group (group C): add 100 μl of 1640 culture medium with 5% calf serum to each well;

[0031] (2) Negative control group: 1640 culture solution added with 5% calf serum, and rhHGF solvent or an equal dose of lyoprotectant;

[0032] (3) rhHGF group: 100 μg / bottle of rhHGF was prepared in 1640 culture medium with 5% calf serum, and the final concentrations were 0.01, 0.1, 0.2, 0.5, 1.0, 5.0 μg / ml.

[0033] 8 wells per group, set at 37°C, 5% CO 2 After culturing ...

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Abstract

The method of detecting the bioactivity of recombinant human liver cell growth factor adopts the technological scheme of applying NIH3T3 fibrocyte strain and mitochondrion dehydrogenase activity method to detect the bioactivity of rhHGF product. The method is one simple, feasible, specific and sensitive rhHGF bioactivity detecting method.

Description

Technical field: [0001] The invention relates to a detection method of biological activity, more specifically a detection method of recombinant human hepatocyte growth factor (rhHGP) biological activity. Background technique: [0002] As one of the most important organs of the human body, the liver bears the heavy responsibility of the "chemical factory" of the human body, and its damage will cause a series of effects on the body. my country is a high-incidence area of ​​viral hepatitis, with more than 30 million active patients. How to prevent hepatitis liver damage and block the development chain of "hepatitis-cirrhosis-liver cancer" is a major issue for medical basic and clinical workers. [0003] Promoting liver cell regeneration is the key to prevent and treat liver damage, and liver cell regeneration is regulated by a variety of cytokines, among which rhHGF is an important factor that promotes liver cell proliferation and repairs liver damage. The study of rhHGF has b...

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Application Information

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IPC IPC(8): C12Q1/32
Inventor 李茹冰王妍柴向东万华印孔祥平
Owner SHENZHEN NEPTUNUS INTERLONG BIO TECHN HLDG
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