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A kind of chicken alpha interferon and preparation method thereof

A technology of chicken interferon and interferon, applied in the field of veterinary biological products, can solve the problems of complex product components, high price, and virus pollution

Active Publication Date: 2021-07-02
DALIAN SANYI ANIMAL MEDICINE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The interferon products produced by the induction agent to induce immune cells to extract interferon protein face two problems: one is that a sufficient number of peripheral blood leukocytes must be ensured in mass production, and the other is that there is potential virus contamination with viruses as the induction agent
In addition, the yield of interferon extracted by this method is limited, the product composition is complex, and the price is expensive, which limits its clinical application

Method used

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  • A kind of chicken alpha interferon and preparation method thereof
  • A kind of chicken alpha interferon and preparation method thereof
  • A kind of chicken alpha interferon and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] ——Preparation of chIFN-α

[0042] 1. The production strain is the recombinant Escherichia coli chIFN-α-pBV220 strain constructed by the inventor from the pBV220 temperature-controlled expression vector with the preservation number CGMCCNo.13431 as the production strain (the strain was delivered on December 07, 2016 Beijing Chaoyang District, Beichen West Road No. 1, No. 3, Institute of Microbiology, Chinese Academy of Sciences, China Microbiology Collection Management Committee General Microbiology Center CGMCC No.13431), and successfully induced the expression of biologically active chIFN-α protein by adjusting the culture temperature.

[0043] 2. Composition of culture medium

[0044] (1) SOB liquid medium (SOB+Amp + ): Soybean peptone 40.0g, yeast powder 10.0g, sodium chloride 0.60g, potassium chloride 0.20g, appropriate volume of purified water to dissolve the above ingredients, add purified water to 1000ml, adjust the pH value to 7.0, autoclave . After the cultu...

Embodiment 2

[0057] ——Determination of biological activity of recombinant chIFN-α protein

[0058] The recombinant chIFN-α Escherichia coli strain can express chIFN-α protein under the condition of 42°C, and the present invention adopts the classic cytopathic inhibition method to detect the biological activity of the target protein expressed by the recombinant chIFN-α-pBV220 strain. The results showed that the chIFN-α protein expressed by the recombinant strain could significantly inhibit the VSV infection of Wish cells, and the results are shown in Table 2-4. According to the data in Table 2, the interferon biological activity was calculated according to the Reed-Muench method. The value of the half cell protection dilution of interferon was calculated as 4 -5.6 , indicating that the 100-fold dilution of the crudely extracted chicken alpha interferon sample was diluted to 4 -5.6 , it can protect half of the cells from damage. Interferon titer = 100 x (5.6 log4 against antilog10). By s...

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Abstract

The invention relates to a chicken alpha interferon and a preparation method thereof. The present invention selects the pBV220 temperature-controlled expression vector to construct recombinant Escherichia coli chIFN-α-pBV220 strain, and the preservation code is CGMCC No.13431. The recombinant strain is used as a production strain to successfully induce and express a large amount of biologically active chIFN-α protein by adjusting the culture temperature. The biologically active chIFN-α protein expressed by the recombinant strain can not only improve the benefit of large-scale industrial production, but also provide a new product for the treatment and prevention of chicken viral diseases.

Description

technical field [0001] The invention relates to a chicken alpha interferon and a preparation method thereof. It belongs to the field of veterinary biological products. Background technique [0002] Chicken viral disease is one of the most serious diseases that endanger the chicken industry. At present, the prevention and control of chicken viral diseases in my country mainly relies on vaccine immunization. Due to the single serotype of vaccine immunization and the rapid mutation of virus strains, vaccine immunization often fails. As a multifunctional cytokine, chicken alpha interferon (chIFN-α) has a wide range of physiological functions such as anti-virus, anti-tumor and immune regulation. , infectious laryngotracheitis and infectious bronchitis have good therapeutic effects, and can significantly improve the immune efficacy of the vaccine. In addition, compared with antibiotics, interferon has the advantages of no toxic side effects and no drug residues. Therefore, the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/56C12P21/02C12R1/19
CPCC07K14/56
Inventor 江国托李凤华潘珊徐凤平
Owner DALIAN SANYI ANIMAL MEDICINE CO LTD