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Process for preparing anti-hepatitis B virus specific transfer facor by gene engineering process

A technology of transfer factor and genetic engineering, which is applied in the field of preparing anti-HBV-specific transfer factor, can solve the problems of high cost, limited source of raw materials, low yield of anti-HBV-specific transfer factor, etc., and achieve low production cost and large output Effect

Inactive Publication Date: 2005-10-19
扬州同人生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its weak point is: because the output of the anti-hepatitis B virus specific transfer factor prepared by this method is low, the cost is high, and the source of raw materials is limited, it is difficult to realize industrialized mass production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Prepare and prepare anti-hepatitis B virus specific transfer factor according to the following steps:

[0014] 1. Cloning of mouse anti-HBV-specific transfer factor gene by mRNA differential display method

[0015] 1. Artificial infection

[0016] Take three healthy adult female BALBc mice of the same age and similar body weight, one of which is artificially infected with hepatitis B virus (subcutaneous injection), one is infected with human herpes virus, and the other is used as a normal control.

[0017] Use commercially available hepatitis B virus surface antigen detection standard kits and herpes virus detection standard kits to detect the production of corresponding antibodies in the mouse serum, and confirm that the artificial infection is successful if positive.

[0018] 2. mRNA extraction from mouse spleen tissue

[0019] Three mice were killed by dislocation of the cervical vertebrae, and the spleen tissues were taken out, rinsed with DEPC-treated saline, and...

Embodiment 2

[0046] 1. Cloning of mouse anti-HBV-specific transfer factor gene by mRNA differential display method

[0047] Healthy adult female BALBc mice were artificially infected with hepatitis B virus. After the infection was successful, the spleen tissue was taken out, rinsed with DEPC-treated saline, and quickly put into liquid nitrogen. After freezing completely, the spleen tissue mRNA could be extracted; RNA was extracted using the Promega RNAgents Total RNA Extraction System (commercially available). Spleen tissue was taken in a denaturant to homogenize the tissue, and extracted with acid-balanced phenol: chloroform: isoamyl alcohol = 125: 24: 1. RNA is detached from DNA and protein, chromatographed from the mixed solution to the aqueous phase, and then the RNA is precipitated and concentrated with isopropanol; the obtained RNA is treated with 1-2 units of DNase without RNase contamination, and then heated at 60°C for 12 minutes , to inactivate DNase, dissolve mRNA in double-dist...

Embodiment 3

[0053] 1. Cloning of mouse anti-HBV-specific transfer factor gene by mRNA differential display method

[0054] A number of healthy adult female BALBc mice were artificially infected with hepatitis B virus. After the infection was successful, the spleen tissue was taken out, rinsed with DEPC-treated saline, and quickly put into liquid nitrogen. After freezing completely, the spleen tissue mRNA could be extracted; spleen tissue Total RNA was extracted using the PromegaRNAgents Total RNA Extraction System. The spleen tissue was homogenized in a denaturing agent, and extracted with acid-balanced phenol: chloroform: isoamyl alcohol = 125:24:1 to separate the RNA from DNA and Protein, from the mixed solution to the aqueous phase, and then use isopropanol to concentrate the RNA precipitation; the obtained RNA is treated with 1-2 units of DNase without RNase contamination, and then heated at 70°C for 8 minutes to make the DNase Inactivation, the mRNA was dissolved in double distilled ...

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Abstract

The gene engineering process for preparing specific transfer factor resisting hepatitis B virus includes the first cloning mouse's specific transfer factor gene resisting hepatitis B virus in mRNA difference showing process; the subsequent constituting recombinant saccharomycete expressing specific transfer factor resisting hepatitis B virus and culturing for large scale expression of specific transfer factor resisting hepatitis B virus; and final crushing, separating and purifying to obtain specific transfer factor resisting hepatitis B virus. The present invention is suitable for large scale production in low cost and great yield and makes it possible to prepare medicine and health article resisting hepatitis B virus.

Description

technical field [0001] The invention relates to a method for preparing anti-hepatitis B virus specific transfer factor. Background technique [0002] Anti-hepatitis B virus-specific transfer factor is a biological product used in the manufacture of drugs for the treatment of hepatitis B. Research has found that when the mother is infected with hepatitis B virus, the placenta contains anti-hepatitis B virus-specific transfer factor, which can be obtained from the placenta The anti-hepatitis B virus-specific transfer factor is extracted. When extracting, the placenta is broken, centrifuged, and the supernatant is taken, which is sterilized and made into a product. Its disadvantages are: due to the low yield and high cost of the anti-hepatitis B virus-specific transfer factor prepared by this method, and limited sources of raw materials, it is difficult to realize industrialized mass production. Contents of the invention [0003] The purpose of the present invention is to pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/09
Inventor 叶满红郭金钢
Owner 扬州同人生物工程有限公司
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