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Dual-target CAR vector combining EpCAM and MSLN single-chain antibody, construction method of dual-target CAR vector and application of dual-target CAR vector in breast cancer

A single-chain antibody and construction method technology, applied in the field of medicine and biology, can solve the problems of lack of specific recognition of tumor-associated antigens, and achieve the effects of avoiding down-regulation of MHC expression, reducing risks, and improving killing efficiency

Inactive Publication Date: 2019-04-09
SHANGHAI YIHAO BIOTECH CO LTD
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Problems solved by technology

[0003] LAK cells, DCs, and CIK cells have all been used as immunotherapy methods for tumors, but all of the above treatments lack the ability to specifically recognize tumor associated antigen (TAA) and kill specific tumor target cells; while tumor immunoediting The process will reduce the expression of the main histocompatibility complex (MHC) on the surface of the target cell on the surface of the tumor cell, forming immune escape, so that the tumor cell can successfully evade the attack of T cells

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  • Dual-target CAR vector combining EpCAM and MSLN single-chain antibody, construction method of dual-target CAR vector and application of dual-target CAR vector in breast cancer
  • Dual-target CAR vector combining EpCAM and MSLN single-chain antibody, construction method of dual-target CAR vector and application of dual-target CAR vector in breast cancer
  • Dual-target CAR vector combining EpCAM and MSLN single-chain antibody, construction method of dual-target CAR vector and application of dual-target CAR vector in breast cancer

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Embodiment Construction

[0036] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be described in further detail below in conjunction with specific embodiments and with reference to the accompanying drawings.

[0037]The present invention provides a dual-target CAR carrier combined with EpCAM and MSLN single-chain antibody. The specific scheme is as follows: comprising a chimeric antigen receptor and a carrier, the chimeric antigen receptor is connected to the carrier, and the chimeric antigen receptor includes two The species-specific structures are respectively: EpCAM single-chain antibody and MSLN single-chain antibody, the nucleotide sequence of the EpCAM single-chain antibody is shown in SEQ ID NO.2, and the nucleotide sequence of the MSLN single-chain antibody is shown in SEQ ID NO. Shown in NO.7. The structural composition of the chimeric antigen receptor is CD8leader-EpCAM scFv-CD8α-CD28-CD137-IRES-CD8leader-MSLN scFv-CD...

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Abstract

The invention belongs to the field of medical biology, and particularly relates to a dual-target CAR vector combining EpCAM and MSLN single-chain antibodies, a construction method of the dual-target CAR vector and an application of the dual-target CAR vector in breast cancer. The dual-target CAR-T therapeutic vector is combined with the EpCAM single chain antibody and the MSLN single chain antibody, specifically CD8leader-EpCAM scFv-CD8 alpha-CD28-CD137-IRES-CD8leader-MSLN scFv-CD8alpha-CD3zeta. The CAR-T cell (Chimeric Antigen Receptor T-Cell) containing the therapeutic vector is obtained byviral infection, and by expressing the CAR structure, targeted identification and killing of breast cancer cells with high expression of EpCAM and MSLN tumor-associated antigens can be achieved.

Description

technical field [0001] The invention belongs to the field of medical biology, and in particular relates to a dual-target CAR carrier combined with EpCAM and MSLN single-chain antibody, a construction method thereof, and an application in breast cancer. Background technique [0002] Breast cancer is one of the common malignant tumors of female reproductive organs, its mortality rate accounts for the first of female malignant tumors, and it is the leading cause of death of female cancer patients. The current treatment methods for breast cancer include surgery, chemotherapy, traditional Chinese medicine, and immunotherapy. In recent years, with the continuous development of basic scientific research, immunotherapy has attracted more and more attention. [0003] LAK cells, DCs, and CIK cells have all been used as immunotherapy methods for tumors, but all of the above treatments lack the ability to specifically recognize tumor-associated antigens (tumor associated antigen, TAA) ...

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Application Information

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IPC IPC(8): C12N15/867C12N15/13C12N15/66C12N5/10C07K19/00A61K35/17A61P35/00
CPCC12N15/86A61K35/17A61P35/00C07K14/7051C07K16/30C07K2319/33C12N15/66C12N2510/00C12N2740/15043
Inventor 田晓丽
Owner SHANGHAI YIHAO BIOTECH CO LTD
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