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Preparation method of tumor vaccine and tumor vaccine prepared by using the method

A tumor vaccine and tumor antigen technology, applied in the field of cellular immunotherapy, can solve the problems of low immune response level and high adjuvant dependence of tumor vaccines, achieve strong immune stimulating activity, improve screening accuracy, and improve efficiency

Pending Publication Date: 2019-11-19
北京微九九科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is to provide a method for preparing a tumor antigen and a tumor antigen prepared by using the method to solve the problem of low immune response level of tumor vaccines in the prior art and high dependence on adjuvants

Method used

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  • Preparation method of tumor vaccine and tumor vaccine prepared by using the method
  • Preparation method of tumor vaccine and tumor vaccine prepared by using the method
  • Preparation method of tumor vaccine and tumor vaccine prepared by using the method

Examples

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preparation example Construction

[0054] The invention discloses a method for preparing a tumor antigen and the tumor antigen prepared by using the method. Those skilled in the art can learn from the content of this article and appropriately improve the process parameters to realize it. It needs to be pointed out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention, and relevant personnel can obviously make changes without departing from the content, spirit and scope of the present invention. Changes or appropriate changes and combinations are made to the content described herein to realize and apply the technology of the present invention.

[0055] In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art.

Embodiment 1

[0057] Example 1: Design of the vaccine

[0058] The tumor tissue and peripheral blood of tumor patients were collected, exome DNA sequencing was performed on tumor tissue and peripheral blood leukocytes, and RNA sequencing was performed on tumor tissue. Sequencing data is analyzed to identify tumor-specific non-synonymous mutations. Antigen prediction software NetMHCpan 4.0 and NetMHCIIpan 3.2 were used to predict the affinities of neoantigens containing non-synonymous mutations to MHCI and MHCII respectively. The stronger the binding ability. Select neoantigen peptides with strong affinity and significantly higher than wild-type antigen peptides as candidate tumor neoantigen peptides, which can effectively stimulate the immune response and have little effect on normal cells expressing wild-type antigen peptides , the candidate tumor neoantigen peptides are shown in Table 1.

[0059] Table 1 Candidate tumor neoantigen peptides

[0060]

Embodiment 2

[0061] Example 2: Construction and expression of vaccines

[0062] Synthesize the DNA sequence corresponding to the neoantigen peptide and clone it into the pShuttle-CMV plasmid, such as figure 1 As shown, pShuttle-CMV-neo was obtained, and finally verified by DNA sequencing. Transfer pShuttle-CMV-neo into BJ5183 competent cells containing pAdEasy, pick a single clone and identify it, obtain the recombinant adenovirus plasmid pAdEasy-pShuttle-CMV-neo that restores the new antigen peptide, and construct the empty recombinant adenovirus plasmid pAdEasy at the same time -pShuttle-CMV. HEK293 cells were transfected to obtain recombinant adenovirus Ad-neo and empty recombinant adenovirus Ad-Vec.

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Abstract

The invention discloses a preparation method of a tumor vaccine. The method includes: cloning a gene sequence of a tumor antigen obtained by screening into a virus expression vector to prepare a recombinant virus to obtain the tumor vaccine; inserting a signal peptide sequence shown in SEQ ID No. 1 or a sequence having a homology of more than 80% with the signal peptide sequence to 5 'end of the gene sequence of the first tumor antigen, and inserting an MHC transport signal sequence shown in SEQ ID No. 2 or a sequence having more than 80% homology with the MHC transport signal sequence to 3 'end of the gene sequence of the last tumor antigen; wherein the virus expression vector is an adenovirus, a retrovirus or a lentivirus, preferably the adenovirus. According to the method, the tumor antigen is screened by a specific method, the virus vector, particularly the adenovirus vector, is used for expressing and preparing the vaccine for the tumor antigen. The vaccine has strong immune stimulation activity and can reduce the requirement on immune adjuvants.

Description

technical field [0001] The invention relates to the technical field of cellular immunotherapy, in particular to a method for preparing a tumor vaccine and a tumor vaccine prepared by using the method. Background technique [0002] Tumor cells are characterized by genome instability and rapid accumulation of somatic mutations. Genome sequencing shows that tumor cells are highly heterogeneous, with hundreds or thousands of somatic mutations in cancer patients, and the types of mutations are also diverse , including point mutations, insertion-deletion mutations, gene amplification, gene rearrangement, etc. Some mutations can cause mutations in the protein encoded by the gene, and the mutated protein can lead to uncontrolled cell proliferation, leading to tumorigenesis. [0003] Cancer vaccines have always been a research hotspot in the field of tumor treatment, but there are many difficulties in the research process, such as the selection of antigens, adjuvants, and delivery m...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00C12N15/12C12N15/861
CPCA61K39/0011A61P35/00C12N15/86C07K14/4748A61K2039/80C12N2710/10043Y02A50/30
Inventor 聂凌虎池振奋何佳
Owner 北京微九九科技有限公司
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