Single-chain antibody targeting ROR1, chimeric antigen receptor T cell, preparation method and application thereof

A chimeric antigen receptor and single-chain antibody technology, applied in the field of medical biology, can solve problems such as limitations, and achieve high specificity, strong affinity activity, internalization activity, and good stability.

CN109836498AInactive Publication Date: 2019-06-04SHENZHEN BINDEBIOTECH CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2019-06-04
Estimated Expiration
Not applicable · inactive patent

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Abstract

The present invention provides a single-chain antibody targeting ROR1, the single-chain antibody targeting ROR1 comprises an amino acid sequence shown in SEQ ID NO: 1. The present invention also provides a chimeric antigen receptor T cell comprising the single-chain antibody targeting ROR1, wherein the chimeric antigen receptor targeting ROR1 can specifically target ROR1, the cellular immunity effect of T cells can be activated, the ROR1 positive tumor cells can be efficiently and specifically killed, the chimeric antigen receptor has lasting cell vitality and lethality, and damage is hardly generated on the normal cells. The invention also provides a preparation method and an application of the chimeric antigen receptor T cell targeting ROR1.
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Description

technical field

[0001] The invention relates to the field of medical biology, in particular to a ROR1-targeting single-chain antibody, chimeric antigen receptor T cells, and a preparation method and application thereof. Background technique

[0002] Malignant tumors are the most serious diseases that endanger human health. Every year, about 3 million people in China are diagnosed with new tumors, and 2 million people die of cancer. Among them, the morbidity and mortality of lung cancer are increasing the fastest, and it is one of the malignant tumors that threaten the health and life of the population the most. Recent studies have shown that the tyrosine protein kinase transmembrane receptor (Receptortyrosine kinase like orphan receptor 1, ROR1) is expressed in a variety of B cell malignancies, and subsequently in lung cancer, mantle cell lymphoma (MCL), ROR1 has been identified on the surface of a number of cancers including subgroups of acute lymphoblastic leukemia (ALL),...

Examples

Embodiment 1

[0079]A method for preparing a chimeric antigen receptor T cell targeting ROR1, comprising the following steps:

[0080] (1) Prepare the gene sequence of chimeric antigen receptor CAR-ROR1 targeting ROR1

[0081] The coding genes of the signal peptide, the single-chain antibody targeting ROR1, the CD8α hinge region, the CD8 transmembrane region, the 4-1BB signal region and the CD3ζ signal region are respectively prepared, and the coding genes of the signal peptide are shown in SEQ ID NO: 15 The coding gene of the single-chain antibody targeting ROR1 is shown in SEQ ID NO: 2, the coding gene of the CD8α hinge region is shown in SEQ ID NO: 7, and the coding gene of the CD8 transmembrane region is shown in SEQ ID As shown in NO: 9, the gene encoding the 4-1BB signal region is shown in SEQ ID NO: 11, and the gene encoding the CD3ζ signal region is shown in SEQ ID NO: 13.

[0082] The genes encoding the above signal peptide, ROR1-targeting single-chain antibody, CD8α hinge region,...