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A combination of lung cancer antigens and its application, cytotoxic T lymphocytes

A lymphocyte and cytotoxic technology, applied in the field of biomedicine, can solve the problems of inaccurate results, separation from experiments, and high incidence of lung cancer

Active Publication Date: 2020-04-21
BEIJING DCTY BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The incidence of lung cancer is the highest in my country, and the mortality rate is high. With the promotion of precision therapy, conventional chemotherapy, radiotherapy and targeting can no longer meet the individuality and variability of tumors. Therefore, it is necessary to find personalized tumor neoantigens and cultivate individualized CTLs (cytotoxic T lymphocytes) that transform tumor neoantigens will be an effective method for completely eradicating tumors
[0003] The key to the cultivation of CTLs targeting tumor neoantigens is how to select tumor neoantigens. At present, the commonly used method is to predict the affinity between tumor neoantigens and HLA through bioinformatics methods, and select those with high affinity as alternatives. The advantage of tumor neoantigens is fast and high-throughput. The disadvantage is that the results are inaccurate based on the algorithm-based separation experiment. Another classic method is to detect the affinity of tumor neoantigens and HLA tetramers in vitro, and then judge the tumor The effectiveness of the neoantigen, the advantage is that based on the affinity of the test, the result is more accurate, the disadvantage is that the cost is high, it takes a long time, and it is detached from the cell, the effectiveness needs to be further verified

Method used

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  • A combination of lung cancer antigens and its application, cytotoxic T lymphocytes
  • A combination of lung cancer antigens and its application, cytotoxic T lymphocytes
  • A combination of lung cancer antigens and its application, cytotoxic T lymphocytes

Examples

Experimental program
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Effect test

Embodiment 1

[0040] 1. Lung cancer-associated neoantigens:

[0041] The whole exome sequencing results of patient Li’s tumor tissue were analyzed for tumor neoantigens (Dingcheng Peptide Source Bio-Information Technology Co., Ltd.), and 80 antigens were synthesized (Nanjing GenScript Polypeptide Synthesis Department);

[0042] Table 1 Peptide sequence and corresponding number

[0043]

[0044]

[0045]

[0046]

[0047] 2. Preparation of APC by co-culture:

[0048] 1) Resuscitated frozen PBMC (2.5×10 7 cells), dilute the freezing solution with 1640+10% FBS, 1500rpm for 5min, resuspend with 10mL1640+10%FBS, 1×10 6 cells / mL, 37°C, 5% CO 2 Recovery culture for 24h;

[0049] 2) Prepare the peptide, 1640+10% FBS+200U / mL IL-2+1600U / mL GM-CSF, the final concentration of the peptide is 50ng / mL, mix well and set aside;

[0050] 3) Collect the cells by centrifugation at 1500rpm for 5min, resuspend the cells with peptide solution, and adjust the density to 1×10 6 cells / mL;

[0051]...

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Abstract

The invention provides a lung cancer antigen combination and an application thereof, and cytotoxic T lymphocytes, and belongs to the technical field of biological medicines. The amino acid sequences of lung cancer antigens are as shown in SEQ ID NO.1-10. The lung cancer antigen combination is used for co-culturing peripheral blood mononuclear cells; and the obtained cytotoxic T lymphocytes can effectively kill cells expressing lung cancer associated antigens.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a lung cancer antigen combination and application thereof, and cytotoxic T lymphocytes. Background technique [0002] The incidence of lung cancer is the highest in my country, and the mortality rate is high. With the promotion of precision therapy, conventional chemotherapy, radiotherapy and targeting can no longer meet the individuality and variability of tumors. Therefore, it is necessary to find personalized tumor neoantigens and cultivate individualized CTLs (cytotoxic T lymphocytes) that transform tumor neoantigens will be an effective method for completely eradicating tumors. [0003] The key to the cultivation of CTLs targeting tumor neoantigens is how to select tumor neoantigens. At present, the commonly used method is to predict the affinity between tumor neoantigens and HLA through bioinformatics methods, and select those with high affinity as alternativ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N5/0783A61K39/00A61K35/17A61P35/00
CPCA61K35/17A61K39/0011A61K2039/86A61P35/00C07K14/4748C12N5/0638C12N2502/11
Inventor 焦顺昌张嵘周子珊王海燕
Owner BEIJING DCTY BIOTECH CO LTD
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