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Chimeric antigen receptors targeting CEA

A chimeric antigen receptor and antigen technology, which is applied in the field of biomedicine, can solve the problems of weak antigenicity, high mortality of tumor patients, and inability to effectively activate specific immune responses, etc., and achieves small side effects, high activity, and improved killing Effects and staying power

Active Publication Date: 2020-08-21
北京百替生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the weak antigenicity of CEA, it cannot effectively activate the body's specific immune response, allowing tumor cells to escape immune killing, resulting in a high mortality rate for patients with this type of tumor
So far, there are still few CAR-T technologies for detecting CEA targets in the existing technology, and CAR still has a lot to do in improving the sensitivity and specificity of binding to CEA targets, and better activating T cells technology space

Method used

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  • Chimeric antigen receptors targeting CEA
  • Chimeric antigen receptors targeting CEA
  • Chimeric antigen receptors targeting CEA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Preparation of embodiment 1 sc-Fv

[0040] 1. Animal immunization and preparation of monoclonal antibodies

[0041] The polypeptide fragment of the extracellular domain of human CEA protein was used as the immunogen, mixed with Freund's complete adjuvant, and immunized 6-week-old female Balb / c mice (the dosage of the immunogen was 100 μg / mouse), and the immunization method was multi-subcutaneous Spot immunization; two weeks later, the immunogen was mixed with incomplete Freund's adjuvant, and the mice were boosted (the dosage of immunogen was 50 μg / mouse), and the immunization method was subcutaneous multi-point immunization; every two weeks Booster immunization was carried out in the same way, a total of 3 booster immunizations. On the 7th day after the last booster immunization, the eyeballs of the mice were removed, blood was collected from the orbital venous plexus of the mice, and the serum was separated by centrifugation, and the antibody titer of the serum was d...

Embodiment 2

[0061] Example 2 Construction of Chimeric Antigen Receptor

[0062] Signal peptide, CEA antigen-binding domain (sc-Fv prepared in Example 1), IL-10R1 ectodomain, IL-15Rβ chain, CD28 co-stimulatory domain, CD3ζ T cell signaling region through whole gene synthesis:

[0063] CEA-CAR: signal peptide-CEA scFv-IL-10R1-IL-15Rβ-CD28-CD3ζ;

[0064] The gene sequence number of CD28 is XM_006712862.1; the gene sequence number of CD3ζ is AF228312.1; the sequence selection refers to the structure of the chimeric antigen receptor designed by Sadelain M, Nature biotechnology, 2013, 31(1):71-5.

Embodiment 3

[0065] Example 3 Construction of CEA-CAR-T lentivirus

[0066] Cloning of CEA-CAR fragments into lentiviral vector pc DNA TM For 3.1(+), the selected insertion site is NheI / BamHI. The sequence was provided by the applicant, and the assembly procedure was completed by Suzhou Gemma Gene Co., Ltd.

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Abstract

The invention relates to the field of biological medicine, in particular to a chimeric antigen receptor taking CEA as a target. The chimeric antigen receptor comprises an antigen binding region, an IL-10R1 extracellular domain, an IL-15RBeta chain, a costimulatory structure domain and a functional structure domain of a T cell signaling region in sequence.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a chimeric antigen receptor targeting CEA. Background technique [0002] CAR-T (Chimeric Antigen Receptor T-Cell Immunotherapy), that is, Chimeric Antigen Receptor T-Cell Immunotherapy. This therapy is a new type of cell therapy that has been around for many years but has only been improved and used clinically in recent years. It has remarkable efficacy in the treatment of acute leukemia and non-Hodgkin's lymphoma, and is considered to be one of the most promising tumor treatment methods. Like all technologies, CAR-T technology has also undergone a long evolutionary process. It is during this series of evolutionary processes that CAR-T technology has gradually matured. [0003] The key to this new therapeutic strategy is artificial receptors known as chimeric antigen receptors (CARs) that recognize target cells and, when genetically modified, express the CARs on the patient's T cells...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/867C12N5/10A61K35/17A61P35/00
CPCA61K35/17A61P35/00C07K14/5428C07K14/5443C07K16/3007C07K2317/565C07K2319/00C12N15/86C12N2740/15043
Inventor 郭磊
Owner 北京百替生物技术有限公司
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