Double-target CAR-T therapy vector for colon cancer, and construction method and application of double-target CAR-T therapy vector

A colon cancer, dual-target technology, used in applications, chemical instruments and methods, botanical equipment and methods, etc., to achieve good therapeutic effects, inhibit release, and reduce damage.

Inactive Publication Date: 2018-10-12
SHANGHAI YIHAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The above patents are all related research on CAR-T cell therapy, but there is currently a lack of vectors or related research on dual-target CAR-T therapy for colon cancer

Method used

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  • Double-target CAR-T therapy vector for colon cancer, and construction method and application of double-target CAR-T therapy vector
  • Double-target CAR-T therapy vector for colon cancer, and construction method and application of double-target CAR-T therapy vector
  • Double-target CAR-T therapy vector for colon cancer, and construction method and application of double-target CAR-T therapy vector

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Embodiment

[0084] (1) Plasmid extraction

[0085] Preparation method of LB liquid medium: Weigh 5g liquid medium dry powder into a 500mL Erlenmeyer flask with an electronic balance, add 100mL ultrapure water, seal with tin foil, sterilize in a high-pressure steam sterilizer, and cool to 40°C~ At 50°C, add 0.2% ampicillin at a ratio of 1000:1, mix carefully, transfer to a clean 500mL reagent bottle for later use, and store at 4°C.

[0086] The preparation method of LB solid medium: Weigh 5g solid medium dry powder into a 500mL Erlenmeyer flask with an electronic balance, add 100mL ultrapure water, seal it with tin foil, sterilize it in a high-pressure steam sterilizer, and cool it to 40°C~ At 50°C, add 0.2% ampicillin at a ratio of 1000:1, mix carefully, invert the plate, and after solidification, stick a parafilm and store at 4°C.

[0087] Take out the glycerol bacteria from the -80°C refrigerator. The glycerol bacteria contain lentiviral expression plasmids pCDH-EF1α-MCS-(PGK-Puro) and...

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Abstract

The invention relates to a double-target CAR-T therapy vector for colon cancer, and a construction method and application of the double-target CAR-T therapy vector. The double-target CAR-T therapy vector consists of two parts comprising a lentiviral expression vector pCDH-EF1aplpha-MCS-(PGK-Puro) and a CAR structure. The specific structure of the CAR is Igkappa-iRGD-CEAscFv-(G<4>S)<5>-Her-2scFv-CD8alpha-CD28-CD137-CD3zeta-T2A-CCL19. A T cell containing the therapy vector, namlely the CAR-T cell (Chimeric Antigen Receptor T-Cell), is obtained through the mode of virus infection, and the CAR-T cell can be used for targeted identification and killing of colon cancer cells by expressing the CAR structure.

Description

technical field [0001] The present invention relates to a CAR-T therapeutic vector, in particular to a dual-target CAR-T therapeutic vector for colon cancer and its construction method and application. Background technique [0002] Colon cancer is a malignant lesion of the colonic mucosal epithelium under the action of various carcinogenic factors such as environment or heredity, and is one of the common malignant tumors. In recent years, the morbidity and mortality in our country are on the rise. At present, the main treatment methods include surgery, chemotherapy, radiotherapy, and traditional Chinese medicine. However, the therapeutic effect of the above methods is limited, and the side effects are relatively large. Although the multidisciplinary team (MDT) diagnosis and treatment model based on surgery for colon cancer and immunotherapy are gradually playing an increasingly important role, they cannot achieve satisfactory results. The ability to specifically recognize...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/867C12N5/10C12N5/0783A61K35/17A61P35/00
CPCA61K35/17C07K14/521C07K14/7051C07K16/3046C07K2319/00C12N5/0636C12N15/86C12N2510/00C12N2740/15043
Inventor 田晓丽
Owner SHANGHAI YIHAO BIOTECH CO LTD
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