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CD24 specific antibody and anti-CD24-CAR-T cell

A CD24, antibody technology, applied in the field of adoptive immune gene therapy of tumors, can solve the problems of low safety and high recurrence rate

Active Publication Date: 2018-08-07
GRACELL BIOTECH SHANGHAI CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are still many shortcomings in the current research on chimeric antigen receptors, and there are still problems such as high recurrence rate and low safety.

Method used

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  • CD24 specific antibody and anti-CD24-CAR-T cell
  • CD24 specific antibody and anti-CD24-CAR-T cell
  • CD24 specific antibody and anti-CD24-CAR-T cell

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0113] Antibody preparation

[0114] Any method suitable for producing monoclonal antibodies can be used to produce anti-CD24 antibodies of the invention. For example, animals can be immunized with linked or naturally occurring CD24 homodimers or fragments thereof. Suitable immunization methods may be used, including adjuvants, immunostimulants, repeated booster immunizations, one or more routes may be used.

[0115] Any suitable form of CD24 can be used as an immunogen (antigen) for producing non-human antibodies specific to CD24 and screening the biological activity of the antibodies. The priming immunogen can be full-length mature human CD24, including native homodimers, or single / multiple epitope-containing peptides. Immunogens can be used alone or in combination with one or more immunogenicity enhancers known in the art. Immunogens can be purified from natural sources, or produced in genetically modified cells. The DNA encoding the immunogen can be genomic or non-geno...

Embodiment 1

[0191] Example 1 Production of CAR lentivirus

[0192] Lentiviruses were prepared by the following steps:

[0193] Day 1:

[0194] 1. Place 5×10 6 HEK293FT cells were seeded into 100 mm diameter petri dishes;

[0195] Day 2:

[0196] 2. Check to make sure the cells are 70%-90% confluent;

[0197] 3. Prepare the transfection complex for each 100 mm diameter Petri dish as follows:

[0198] a. In 1.5ml tube A: dilute 2.5μg CAR (chimeric antigen receptor) DNA plasmid (plasmid) and 20μL lentiviral packaging mix (ALSTEM, catalog number VP100; see Appendix B3) to 0.5ml DMEM or Opti- In MEM serum-free medium, mix gently;

[0199] b. In 1.5ml tube B: Dilute 30μL Nanofect Transfection Reagent (ALSTEM, catalog number NF100) into 0.5ml DMEM or Opti-MEM serum-free medium, mix gently;

[0200] c. Add the NanoFect / DMEM in tube B to the DNA / DMEM solution (tube A), vortex for 5-10 seconds, and incubate the DMEM-plasmid-NanoFect mixture at room temperature for 15 minutes;

[0201] 4. Ad...

Embodiment 2

[0212] Example 2 lentiviral packaging system

[0213] Product Description

[0214] Product name: SuperLenti TM Lentivirus Packaging System

[0215] manual:

[0216] For the production of lentiviral particles, three components are generally required: 1) a lentiviral vector containing the foreign gene of interest, 2) a packaging vector containing all the necessary viral structural proteins, 3) expressing vesicular stomatitis virus (VSV) sugars Envelope carrier for protein (G). The third-generation lentiviral packaging system provides maximum biosafety because the lentiviral Rev gene is provided as a separate vector from other structural genes, further eliminating the possibility of reverse recombination of the vector into replication-competent virus particles. Third-generation lentiviral packaging mixes only support lentiviral expression vectors with chimeric 5’LTRs, where the HIV promoter is replaced by CMV or RSV, thus making it independent of TAT.

[0217] The SuperLent...

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PUM

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Abstract

The invention relates to a monoclonal anti-human CD24 antibody, for example, a single-chain variable region fraction which includes a VH having the amino acid represented as the SEQ ID NO.5 and a VL having the amino acid represented as the SEQ ID NO.6; the invention also relates to a chimeric antigen receptor fusion protein, which includes, from a N-terminal to a C-terminal: (i) the single-chain variable region fraction in the invention; (ii) a transmembrane structural domain; (iii) at least one co-stimulus structural domain; and (iv) an activation domain.

Description

technical field [0001] The present invention relates to CD24-specific antibodies and anti-CD24-CAR-T cells, which can be used in the field of adoptive immune gene therapy for tumors. Background technique [0002] Immunotherapy is an emerging and very promising approach to treating cancer. T cells, or T lymphocytes, are the immune system's potent weapons, constantly seeking out foreign antigens and distinguishing normal cells from abnormal cells (cancerous or infected). Genetic modification of T cells with CAR (chimeric antigen receptor) constructs is the most common approach to engineer tumor-specific T cells. The infusion of CAR-T cells targeting tumor-associated antigens (TAAs) into patients (termed adoptive cell transfer or ACT) represents an effective immunotherapy approach. The advantage of CAR-T technology over chemotherapy or antibodies is that the reprogrammed engineered T cells can proliferate and persist in the patient (the "active drug"). [0003] A CAR typical...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C07K19/00C12N15/13C12N15/62C12N5/10A61K35/17A61P35/00A61P35/02
CPCA61K35/17C07K14/7051C07K16/2896C07K2319/33C07K2319/02C07K2317/56C07K2317/622C12N2510/00
Inventor 曹卫
Owner GRACELL BIOTECH SHANGHAI CO LTD
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