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Preparation method of AFP/HBsAg double-targeting Dexosome anti-hepatoma anti-tumor vaccine

An anti-cancer, dual-targeting technology, applied in the field of tumor biological immunotherapy, can solve problems such as unstable biological activity, achieve strong anti-tumor immune effect, and improve the ability of liver cancer antigen recognition.

Inactive Publication Date: 2015-07-01
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

Among these methods, transfection of DC with tumor cell-derived RNA and adenovirus-mediated infection of DC with various tumor-associated antigens are the most effective, but tumor cell-derived RNA has disadvantages such as unstable biological activity.

Method used

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  • Preparation method of AFP/HBsAg double-targeting Dexosome anti-hepatoma anti-tumor vaccine
  • Preparation method of AFP/HBsAg double-targeting Dexosome anti-hepatoma anti-tumor vaccine
  • Preparation method of AFP/HBsAg double-targeting Dexosome anti-hepatoma anti-tumor vaccine

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Embodiment Construction

[0048] Below in conjunction with accompanying drawing and embodiment the content of the present invention is described in detail

[0049] 1. Construction of adenovirus recombinant plasmids (Ad-HBsAg, AFP) containing HBsAg and AFP genes:

[0050] (1) The results of enzyme digestion identification of the target gene show that the plasmid pCR3.1-S was digested with Xbal I and Hind III, and the plasmid pEGFP-AFP was digested with EcoRI and BamH I, and identified by 1% agarose gel electrophoresis, such as figure 1 shown. (2) Preparation of Escherichia coli Competent Cells (CaCl 2 Law)

[0051] I. Take the revived strains of Escherichia coli stored at -70°C, scrape the surface of the frozen culture with a sterilized inoculation needle, inoculate on the surface of the LB culture plate, and incubate at 37°C for 16 hours.

[0052] II. Pick a single colony from the LB plate, inoculate it in 10ml LB liquid medium, and cultivate it with shaking at 200rpm at 37°C for about 12 hours unti...

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Abstract

The invention discloses a preparation method of an AFP / HBsAg double-targeting Dexosome anti-hepatoma anti-tumor vaccine. A novel double-targeting anti-hepatoma anti-tumor vaccine is prepared by the following steps: by using a recombinant human adenovirus vector as one target of a hepatoma associated antigen AFP gene and combining the characteristics of primary hepatic carcinoma of China, combining a specific antigen HBsAg of hepatoma associated virus as an immune immunotherapy target; importing DC; and extracting Dexosome, wherein the anti-hepatoma immunological effect induced is enhanced. The novel Dexosome anti-tumor vaccine can be combined with an antigen peptide at the same time through MHC-I / II molecules, so that not only can the CD8+cytotoxic T cell be activated, but also the CD4+auxiliary T cell can be activated. The method overcomes the problems in the prior at that the immunological effect induced by a single tumor antigen is weak, DC is complex to prepare and the like and is a preparation method of a novel non-cellular Dexosome anti-hepatoma anti-tumor vaccine.

Description

technical field [0001] The invention belongs to the field of tumor biological immunotherapy, and in particular relates to the preparation of AFP / HBsAg double-targeted Dexosome anti-hepatoma vaccine. Background technique [0002] Primary liver cancer (hepatocellular carcinoma, HCC) is one of the most common malignant tumors in my country. Epidemiological data show that the incidence of HCC in the HBsAg-positive population is 12-300 times that of other populations. The carrier rate of HBV in Chinese population is about 10%, and 90% of primary liver cancer patients are found to be HBsAg positive. Although some progress has been made in the treatment of primary liver cancer, the effect on the treatment and prevention of recurrence and metastasis of advanced patients is still not satisfactory. Therefore, it is necessary to explore a new and effective treatment approach. With the discovery and confirmation of human tumor antigens and tumor antigen genes, new methods are provided...

Claims

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Application Information

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IPC IPC(8): C12N15/861C12N15/51C12N15/12C12N5/10A61K39/29A61K39/00A61P31/20A61P35/00A61P1/16
Inventor 杨静悦李宵张红梅刘文超
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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