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Chimeric antigen receptor with stable antigen binding units, method for preparing chimeric antigen receptor and application thereof

A technology for combining units and antigens, applied in chemical instruments and methods, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc.

Active Publication Date: 2015-08-12
GUANGZHOU BIO GENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many unresolved issues before this treatment option can be widely accepted
One of these issues is the stability of single-chain variable fragment (scFv) expressed as a fusion protein on the cell surface

Method used

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  • Chimeric antigen receptor with stable antigen binding units, method for preparing chimeric antigen receptor and application thereof
  • Chimeric antigen receptor with stable antigen binding units, method for preparing chimeric antigen receptor and application thereof
  • Chimeric antigen receptor with stable antigen binding units, method for preparing chimeric antigen receptor and application thereof

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Experimental program
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Effect test

Embodiment Construction

[0030] 1. Lentiviral Packaging

[0031] Lentiviral packaging was performed using a four-plasmid system. The 4 plasmid systems respectively express gag / pol, Rev, VSV-G required for lentiviral vector packaging, and artificial chimeric antigen receptors composed of engineered stable single-chain antibodies. The 4 plasmids were transiently transfected as compared. The total mass is 10 μg. The above plasmid was added to a certain volume of sterile water, then 62.5 μl of 2M CaCl2 solution was added, and finally 500 μl of 2×HBS (pH=7.05) buffer was added. Add the above solution to a 10cm Petri dish covered with cells and mix gently. Cultivate in a 37°C, 5% CO2 cell incubator for 6h. After 6h, replace the fresh medium and continue the culture. And a 10 mM sodium butyrate solution was added. After 49-72 hours, the culture supernatant of the lentivirus was collected for purification and detection.

[0032] 2. Expansion of T cells

[0033]30ml of whole blood was collected from eac...

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Abstract

The invention relates to a chimeric antigen receptor with stable antigen binding units. The chimeric antigen receptor comprises the extracellular antigen binding units, trans-membrane domains and intracellular co-stimulation signal domains. The antigen binding units can be stably expressed as compared with specific tumor targets and comprise heavy chains and light chains which are connected with one another by hinges of SEQ ID NO.2 (sequence identity number.2) amino acid sequence codes. The invention further relates to cells for expressing the receptor and application of the receptor.

Description

technical field [0001] The invention relates to a chimeric antigen receptor containing a highly stable antigen-binding unit, a preparation method thereof, a cell containing the artificial chimeric antigen receptor, and its application. Background technique [0002] Adoptive immunotherapy uses T cells that have not been genetically modified and has already demonstrated its potential in the treatment of certain cancers and viral infections. Examples include graft-versus-leukemia (GVL) elicited by donor lymphatic infusion (DLI) for the treatment of relapsed leukemia and injection of ex vivo expanded tumor-infiltrating lymphocytes (TIL) in combination with pre-treatment lymphodepletion for treatment. Advanced melanoma. However, both DLI and TIL treatments have severe side effects and lack of specificity that limits their efficacy. Attempts have been made to discover potent immunogenic antigens for modification of effector T cells that can selectively target leukemia or other t...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N5/10A61K38/17A61K47/48A61P35/02
Inventor 李胜峰秦超丁雯俞金泉
Owner GUANGZHOU BIO GENE TECH CO LTD
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