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Dual-target tumor vaccine based on tumor endothelium marker-8 gene and preparation method thereof

A technology of tumor vaccines and markers, applied in the field of tumor vaccines, can solve the problems of small polypeptide molecules and weak immunogenicity, and achieve the effects of improving effectiveness and safety, promoting tumor cell apoptosis, and good application prospects

Inactive Publication Date: 2012-05-09
ARMY MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of this, one of the objectives of the present invention is to provide a dual-target tumor vaccine based on the TEM-8 gene, which not only retains the advantages of safety and easy preparation and purification of the polypeptide vaccine, but also overcomes the small polypeptide molecule, weak immunogenicity, It is not easy to be taken up by antigen presenting cells (APC) in vivo, and can stimulate effective specific cytotoxic T lymphocyte (CTL) response in vivo, and at the same time play the role of inhibiting tumor angiogenesis and inducing tumor cell apoptosis; the present invention The second purpose is to provide a method for preparing the dual-target tumor vaccine based on the TEM-8 gene

Method used

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  • Dual-target tumor vaccine based on tumor endothelium marker-8 gene and preparation method thereof
  • Dual-target tumor vaccine based on tumor endothelium marker-8 gene and preparation method thereof
  • Dual-target tumor vaccine based on tumor endothelium marker-8 gene and preparation method thereof

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Embodiment Construction

[0025] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. The experimental method that does not indicate specific conditions in the preferred embodiment is usually according to conventional conditions, such as described in the Molecular Cloning Experiment Guide (Third Edition, J. Sambrook et al., translated by Huang Peitang, etc., Science Press, 2002) conditions, or as recommended by the manufacturer.

[0026] 1. Preparation of tumor vaccines

[0027] 1. Preparation of TEM-8 gene recombinant eukaryotic expression vector pcDNA3.1 / TEM-8

[0028] (1) Cloning of TEM-8 full-length coding gene

[0029] According to the TEM-8 gene sequence with GenBank accession number NM 018153.3, the following PCR primers were designed and synthesized to amplify the full-length coding gene of TEM-8: Primer F 1: 5'-gccacctttgcgaccctcctgagcttagg-3' (SEQ ID No.3); Primer R 1: 5′-tatttccctgcctccattatactgactcaagcag-3′ (...

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Abstract

The invention discloses a dual-target tumor vaccine based on a tumor endothelium marker-8 gene and a preparation method thereof. The tumor vaccine is formed by combining an eukaryon expression vector recombined by the tumor endothelium marker-8 gene with fusogenic peptide obtained by fusing a membrane penetrating peptide HIV-Tat49-57 and a RGD peptide in the manner of non covalent bonds. The preparation method thereof comprises the following steps: respectively preparing the eukaryon expression vector recombined by the tumor endothelium marker-8 genethe and the fusogenic peptide obtained by fusing the membrane penetrating peptide HIV-Tat49-57 and the RGD peptide; and then, polymerizing the eukaryon expression vector and the fusogenic peptide through electrostatic interaction to form a compound. The tumor vaccine not only reserves the advantages of polypeptide vaccine, such as safety, easy preparation and purification and the like, but also overcomes the shortages, such as small polypeptide molecule, weak immunogenicity and difficult absorption by antigen presenting cells and the like. The vaccine can inhibit the tumor angiogenesis as well as induce the apoptosis of tumor cells, thereby greatly improving the effectiveness and the safety of tumor therapy. The vaccine has wide application prospects in the field of tumor therapy.

Description

technical field [0001] The invention relates to a tumor vaccine, in particular to a dual-target tumor vaccine based on tumor endothelial marker-8 (TEM-8) gene, and also relates to a preparation method of the tumor vaccine. Background technique [0002] Tumor is a disease with high morbidity and high mortality that seriously threatens human health. After surgery, radiotherapy and chemotherapy, biological therapy has become an important means of comprehensive treatment of tumors. Currently, tumor biotherapy strategies include targeted therapy of tumor-associated genes or tumor suppressor genes, and immunotherapy of tumor-associated antigens. [0003] Studies have found that more than 90% of human tumors are solid tumors. The growth and metastasis of solid tumors require the formation of new blood vessels. Inhibiting the formation of new blood vessels can effectively inhibit the growth and metastasis of tumors to a certain extent. Therefore, inhibiting tumor angiogenesis has b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61P35/00A61K47/48A61K47/42A61K48/00A61K39/385A61K47/64
Inventor 杨曌吴玉章
Owner ARMY MEDICAL UNIV
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