Application of sequence-optimized secreted form FAPalpha-based tumor DNA vaccine

A DNA vaccine and vaccine technology, applied in the field of tumor DNA vaccine, to achieve the effect of good protein expression level, high safety and strong anti-tumor activity

Pending Publication Date: 2019-09-06
JILIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This shows that the full-length FAPα still has certain safety issues when applied

Method used

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  • Application of sequence-optimized secreted form FAPalpha-based tumor DNA vaccine
  • Application of sequence-optimized secreted form FAPalpha-based tumor DNA vaccine
  • Application of sequence-optimized secreted form FAPalpha-based tumor DNA vaccine

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Experimental program
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Embodiment Construction

[0033] 1. Construction and identification of CpVR-shF(m) and CpVR-OshF(m) DNA vaccines

[0034] The vector used is the eukaryotic expression plasmid VR1012 with the introduction of CpG motif, referred to as CpVR, which contains promoter CMV, intron A, BGH sequence, CpG sequence, kanamycin resistance sequence, a total of 5305bp, see the map Figure 1A .

[0035] Using CpVR-hF(m) (and CpVR-FAP) as a template, the nucleotide sequence SEQ ID NO: 1 was obtained by multi-step PCR, and BglII restriction sites and homologs identical to the sequences at both ends of the vector were introduced at both ends. For the source arm sequence, perform DNA electrophoresis and gel recovery on the target DNA fragment and the vector fragment obtained by digestion with BglII to obtain the target fragment and vector, use a homologous recombination kit, incubate at 50°C for 15 minutes, transform Escherichia coli, and coat the card Namycin-resistant LB plates were incubated overnight at 37°C. Select p...

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Abstract

The invention relates to an application of a sequence-optimized secreted form FAPalpha-based tumor DNA vaccine, which belongs to the field of a tumor DNA vaccine. The invention relates to a FAPalpha-derived secreted form FAPalpha extracellular segment sequence shF(m) and its codon-optimized sequence OshF(m) and an application of a DNA vaccine constructed based on two sequences for preparing a tumor therapeutic vaccine. The application determines, by immunogenicity experiments and anti-tumor experiments, that the above DNA vaccine has stronger antitumor activity than a DNA vaccine having the original FAPalpha sequence. the sequence OshF(m) has only 76% homology with the original sequence, and can better express the target protein, and the application has a better application prospect in thepreparation of various tumor vaccines based on FAPalpha.

Description

technical field [0001] The present invention relates to the field of tumor DNA vaccines, in particular, the present invention relates to the recombinant DNA carrier CpVR-shF(m) targeting FAPα and its codon-optimized recombinant DNA carrier CpVR-OshF(m) in the preparation of anti-tumor vaccines Applications. Background technique [0002] The tumor microenvironment (TME) is the place where tumors depend for survival, providing the necessary nutrients for tumor growth, while isolating tumor cells from immune cells and preventing immune cells from killing tumor cells. Cancer-associated fibroblasts (CAFs) are an important part of the tumor microenvironment and are closely related to tumor growth, invasion, and metastasis. They can secrete soluble cytokines (such as stromal cell-derived factor: SDF) through direct interaction with tumor cells. -1), remodeling extracellular matrix and other means to promote tumor growth. Fibroblast activating protein α (FAPα) is a specific marker...

Claims

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Application Information

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IPC IPC(8): C12N15/62A61K39/00A61P35/00
CPCC12N9/6424A61K39/0011A61P35/00C07K2319/02
Inventor 张海红于湘晖孔维耿飞郭杰
Owner JILIN UNIV
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