Dual-target CAR-T therapeutic vector for breast cancer and construction method and application thereof

A breast cancer, dual-target technology, applied in the field of breast cancer dual-target CAR-T therapy carrier and its construction, to achieve the effect of lasting vitality, eliminating tumor burden, and strong proliferation

Pending Publication Date: 2018-11-23
SHANGHAI YIHAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The above-mentioned 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 breast cancer

Method used

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  • Dual-target CAR-T therapeutic vector for breast cancer and construction method and application thereof
  • Dual-target CAR-T therapeutic vector for breast cancer and construction method and application thereof
  • Dual-target CAR-T therapeutic vector for breast cancer and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0083] (1) Plasmid extraction

[0084] 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.

[0085] 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.

[0086] Take out the glycerol bacteria from the -80°C refrigerator. The glycerol bacteria contain lentiviral expression plasmids pCDH-EF1α-MCS-(PGK-Puro) a...

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Abstract

The invention provides a dual-target CAR-T therapeutic vector for breast cancer and a construction method and application thereof. The dual-target CAR-T therapeutic vector consists of a lentiviral expression vector pCDH-EF1 alpha-MCS-(PGK-Puro) and a CAR structure. The CAR structure is specifically Igkappa-iRGD-mesothelin scFv-(G4S)5-CD133scFv-CD8 alpha-CD28-CD137-CD3 zeta-2A-CCL19. A virus infection mode is adopted to obtain a T cell containing the therapeutic vector, namely a CAR-T cell (Chimeric Antigen Receptor T-Cell), and breast cancer cells can be recognized and killed in a targeted mode through expression of 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 breast cancer and its construction method and application. Background technique [0002] As a common tumor that threatens women's physical and mental health, breast cancer accounts for about 23% of systemic malignant tumors and 14% of mortality. The current treatment methods for breast cancer mainly include surgery, radiotherapy, chemotherapy or combined therapy, but the therapeutic effect is limited; as a new treatment method, gene therapy has not made breakthroughs because the targeted disease-causing genes cannot be determined. Second, tumor heterogeneity and the complexity of the tumor microenvironment mean that the chances of a single approach to cure breast cancer are slim. In today's multidisciplinary comprehensive treatment, new treatment methods need to be proposed urgently. [0003] CAR-T cell therapy is to artificially o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/85C12N15/867C12N5/10A61P35/00
CPCA61P35/00C07K14/521C07K14/7051C07K14/70517C07K14/70521C07K14/70578C07K16/2896C07K16/3015C07K2317/622C07K2319/00C07K2319/03C07K2319/33C07K2319/74C12N5/0636C12N15/85C12N15/86C12N2510/00C12N2740/15043C12N2800/107
Inventor 田晓丽
Owner SHANGHAI YIHAO BIOTECH CO LTD
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