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Anti-PD-L1-CAR-T, and preparation method and application thereof

A PD-L1, anti-pd-l1-car-t technology, applied in the field of Anti-PD-L1-CAR-T and its preparation and application

Active Publication Date: 2017-01-25
SHANGHAI YUYAN BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although some scholars have used CAR-T technology (such as anti Her-2 CAR-T) in combination with immune checkpoint technology (PD-1 antibody) to compare the effect on tumors when used alone, so far, no anti-CAR-T technology has been used. A precedent for constructing PD-L1 gene into CAR gene

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  • Anti-PD-L1-CAR-T, and preparation method and application thereof
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  • Anti-PD-L1-CAR-T, and preparation method and application thereof

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Embodiment Construction

[0024] Hereinafter, the present invention will be further described in conjunction with the embodiments and drawings, and the embodiments are only used for illustration and not for limiting the protection scope of the present invention.

[0025] The design idea of ​​the present invention is as figure 1 As shown, the specific steps are as follows:

[0026] Part 1: PD-L1 gene construction

[0027] The gene synthesis of PD-L1 secreted protein was completed in Suzhou Jinweizhi Company. This gene adds CD33 sequence to the reported PD-L1 gene (Gene ID: 29126) to promote the expression of PD-L1 secreted protein. Its gene sequence is shown in SEQ ID NO:4.

[0028] Part II: Expression and purification of PD-L1 protein

[0029] 1. PD-L1 gene transfection of COS cells (the transfection is divided into 3 groups, one is the transfection group; the other is the negative control group; the other is the blank control group)

[0030] 1. One day before transfection, take 2ml, 0.7×10 6 cos...

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Abstract

The invention relates to anti-PD-L1-CAR-T, and a preparation method and application thereof. A PD-L1 antibody gene is synthesized with gene segments in a hinge region, a transmembrane region and an intracellular signal region of CAR-T, and a constructed carrier is a PCDH chronic virus carrier: pcDH-CNV-MCS-EF1-copGFP. According to the anti-PD-L1-CAR-T, and the preparation method and the application thereof provided by the invention, an anti-PD-L1 gene is constructed into a CAR gene for the first time, and a new idea is provided for tumor treatment.

Description

[0001] [technical field] [0002] The present invention relates to Anti-PD-L1-CAR-T and its preparation method and application. [0003] [Background technique] [0004] Using the body's own immune system to treat tumors, that is, tumor immunotherapy, has developed rapidly in recent years, and its clinical trials have made remarkable progress. It was rated as the first of the top ten scientific breakthroughs in 2013 by Science magazine. Tumor immunotherapy mainly includes non-specific immune stimulation, immune checkpoint antibodies, adoptive cell therapy, monoclonal T cell receptor therapy and tumor therapeutic vaccines. Adoptive T cell therapy is a kind of tumor immunotherapy, which has become the fourth method of treating tumors after surgery, radiotherapy and chemotherapy. The adoptive immune cell therapy of tumors has undergone early lymphokine-activated killer cell (LAK) therapy, cytokine-induced killer cell (CIK, DC-CIK) therapy, NK cell therapy, and tumor infiltrating l...

Claims

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Application Information

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IPC IPC(8): C12N15/62C12N15/867A61K35/17A61P35/00
CPCA61K35/17C07K14/7051C07K14/70521C07K14/70578C07K16/2827C07K2319/33C07K2319/74C12N15/86C12N2740/15043
Inventor 施炜星杨春霞杜盈房永生
Owner SHANGHAI YUYAN BIOTECH CO LTD
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