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A kind of luciferase embedded in antigenic epitope peptide and its construction method and application

An antigenic epitope and luciferase technology, applied in the fields of application, cancer antigenic components, oxidoreductase, etc., can solve the problems of weak immunogenicity, unclear antigenic epitope peptide, and insufficient sensitivity of tumor immunotherapy, etc., and achieve insertion The effect of short sequences and high sensitivity

Active Publication Date: 2021-12-10
GUANGZHOU JENNIO BIOLOGICAL TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The syngeneic transplantation mouse model is a commonly used animal model for the study of tumor immunotherapy, but the original tumor cell line of the syngeneic transplantation mouse model is usually weak in immunogenicity, and the epitope peptide is often not clear, so it is used to evaluate the sensitivity of tumor immunotherapy not enough

Method used

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  • A kind of luciferase embedded in antigenic epitope peptide and its construction method and application
  • A kind of luciferase embedded in antigenic epitope peptide and its construction method and application
  • A kind of luciferase embedded in antigenic epitope peptide and its construction method and application

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Embodiment 1

[0042] Example 1, Construction of Luciferase Embedded in Epitope Peptide

[0043] According to the murine tumor cell line that needs to be modified and its source (or histocompatible) mouse strain, select the exogenous antigen epitope peptide and construct the combined epitope sequence (called CombiEpi). B16 and MC38 cell lines are derived from C57 mice, select OVA257-264 (the amino acid sequence is SIINFEKL, see the UniProt protein database accession number: A0A2H4Y842, was 2-K b presented) and influzenza A NP 366-374 (amino acid sequence is ASNENMETM, see UniProt protein database accession number: P03466, was 2-D b presented), 4T1 and CT26 cell lines are derived from BALB / c mice, you can choose influzenza A HA 532-540 (amino acid sequence is IYSTVASSL, see UniProt protein database accession number: P03452, which is 2-D dpresented) and respiratory syncytial virus M2 82–90 (amino acid sequence is SYIGSINNI, see UniProt protein database accession number: P04545, was 2-K d p...

Embodiment 2

[0048] Embodiment 2, in vivo imaging experiment

[0049] Experiment: 12 6-week-old female BALB / c mice were randomly divided into 4 groups (PBS group, PADRE+CpG group, A+PADRE+CpG group, A+B+PADRE+CpG group), 3 mice in each group, numbered #1 ~#12.

[0050] Subcutaneous immunization three times at intervals of 14 days; PBS group each subcutaneously injected with 100 μl of PBS (number #1 to #3);

[0051] The PADRE+CpG group (number #4~#6) was subcutaneously injected with 25 μg CpG-ODN 1826 and 50 μg CD4+ T cell (i.e. helper T cell) universal activation peptide PADRE (amino acid sequence is AKFVAAWTLKAAA), mixed and dissolved in 100 μl PBS.

[0052] A+PADRE+CpG group (number #7~#9) was subcutaneously injected with 100 μg antigenic peptide A (antigen epitope peptide, SEQ ID NO: 3), 25 μg CpG-ODN 1826 and 50 μg CD4+ T cell universal activation peptide PADRE , mix and dissolve in 100 μl PBS.

[0053] A+B+PADRE+CpG group (number #10~#12) was subcutaneously injected with 50 μg a...

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Abstract

The invention relates to the field of biotechnology, and specifically discloses a luciferase embedded in an antigen epitope peptide, a construction method and application thereof. The antigenic epitope peptide is: at least composed of one amino acid sequence shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4. The invention constructs an immunogenic antigenic epitope peptide or an antigenic combination epitope peptide, and inserts it into luciferase, so that a tumor immune system with high sensitivity (adjustable) and which can be tracked by means of in vivo imaging technology can be constructed. research model.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a luciferase embedded in an antigenic epitope peptide and its construction method and application. Background technique [0002] The syngeneic model is to inoculate tumor cell lines from the same background into immune-competent inbred mice (such as the commonly used C57BL / 6 or BALB / c strains), and the inoculation site is usually subcutaneous ( Convenient for tumor observation and measurement), in situ (simulating tumor microenvironment), tail vein injection (monitoring tumor metastasis). [0003] Tumor immunotherapy is a new tumor prevention and treatment method that eliminates or controls tumors by restoring or enhancing the body's anti-tumor immune response, including tumor immune checkpoint inhibitors (such as CTL-4 inhibitors, PD-1 / PD-L1 inhibitors), Adoptive cellular immunotherapy (such as CAR-T) and other treatment methods that have shown outstanding clinical efficacy, in add...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/77C12N9/02C12N15/53C07K19/00C12N15/85C12N5/10A61K39/00A61P35/00A61K49/00
CPCC07K14/77C12N9/0069C12N15/85C12Y113/12007C12N5/0693A61K39/0011A61P35/00A61K49/0045C07K2319/00C12N2510/00A61K2039/55561
Inventor 胡策谢鹏唐玉巧
Owner GUANGZHOU JENNIO BIOLOGICAL TECH CO LTD
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