CAR-T (chimeric antigen receptor T cell) storage solution

A chimeric antigen receptor and storage solution technology, applied in the field of medical biology, can solve the problems of affecting cell activity and CAR-T cell therapy effect, and achieve the effect of convenient transportation and clinical application

Inactive Publication Date: 2018-12-28
刘劼 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing storage solution is the storage solution of ordinary immune cells, which will affect the

Method used

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  • CAR-T (chimeric antigen receptor T cell) storage solution
  • CAR-T (chimeric antigen receptor T cell) storage solution
  • CAR-T (chimeric antigen receptor T cell) storage solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0071] Construction of viral plasmids: The single-chain antibody sequence that specifically recognizes the CD19 antigen and the intracellular signaling molecules CD3ζ and 4-1BB were concatenated together by PCR to obtain the CD19-CAR fragment. The selected infection method is lentiviral transduction. Finally, the CD19-CAR fragment was cloned into the lentiviral vector. The selected insertion sites were BamH I and Sal I. The CAR19 was constructed based on the main plasmid and identified by sequencing results. .

[0072] Cells were cultured according to conventional cell culture methods at 37°C, 5% CO 2 T lymphocytes extracted from peripheral blood were cultured in 5% inactivated AB-type human plasma Xvivo15 medium; lentiviral packaging cell line 293T was cultured in DMEM medium containing 10% FBS.

[0073] Extraction of lentiviral packaging plasmid and target plasmid Plasmid transformation:

[0074] 1) Take a tube of 100ul competent cells and place them in an ice bath;

[00...

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Abstract

The invention provides a CAR-T (chimeric antigen receptor T cell) storage solution, and belongs to the field of cytobiology. The storage solution is composition of the CAR-T storage solution suitablefor clinical application. Storage conditions are optimized by a variety of active ingredients, and the storage solution is an aqueous solution containing adenosine, sodium chloride, albumin, angiotensin-II, cell-derived short peptide and peptide compounds, an antibody T1, an antibody T2, an antibody T3, antagonist tocilizumab of cytokine interleukin-6 and cytokine interleukin-15. In the storage solution, activity of the CAR-T under the condition of 4-8 DEG C within 2 days keeps 90% or higher, killing efficiency keeps 50% or higher, and the cell phenotype is stable. The storage solution ensuresstable quality of the cells, can realize long-distance transportation of the cells, and lays a foundation for wide application of the clinical cells.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and in particular relates to a storage solution of chimeric antigen receptor modified T cells (CAR-T). Background technique [0002] With the impact of environmental pollution, population aging, unhealthy lifestyle, and psychological and spiritual factors, the incidence of cancer is increasing year by year all over the world. The incidence of cancer in China is severe, and the incidence and mortality continue to rise. , there are about 4.292 million new cancer cases each year, and about 2.814 million cancer deaths. Lung cancer, gastric cancer, colorectal cancer, liver cancer and esophageal cancer are currently the most common types of malignant tumors in China, accounting for 64% of all cancers; breast cancer, cervical cancer, prostate cancer, thyroid cancer and hematological tumors are the fastest growing incidences of several types. Generally speaking, the incidence rate of each tumor var...

Claims

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Application Information

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IPC IPC(8): A01N1/02A61K35/17A61P35/00A61P35/02
CPCA01N1/0226A61K35/17A61P35/00A61P35/02
Inventor 刘劼梁爱斌黄明辉朱益辉
Owner 刘劼
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