CD24-specific antibodies and anti-CD24-CAR-T cells
A CD24 and antibody technology, applied in the field of adoptive immune gene therapy of tumors, can solve the problems of high recurrence rate and low safety
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0113] Preparation of antibodies
[0114] Any method suitable for producing monoclonal antibodies can be used to produce the anti-CD24 antibodies of the invention. For example, animals can be immunized with a linked or naturally occurring CD24 homodimer or fragment thereof. Appropriate methods of immunization can be used, including adjuvants, immunostimulants, repeated booster immunizations, and one or more routes can be used.
[0115] Any suitable form of CD24 can be used as an immunogen (antigen) for generating non-human antibodies specific for CD24 and screening said antibodies for biological activity. The priming immunogen can be full-length mature human CD24, including native homodimers, or peptides containing single / multiple epitopes. The 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 ca...
Embodiment 1
[0191] Example 1 Production of CAR lentivirus
[0192] Lentiviruses are prepared by the following steps:
[0193] Day 1:
[0194] 1. Put 5×10 6 HEK293FT cells were seeded into 100 mm diameter dishes;
[0195] Day 2:
[0196] 2. Check to ensure 70%-90% cell confluence;
[0197] 3. Prepare the transfection complexes for each 100 mm diameter petri dish as follows:
[0198] a. In 1.5 ml tube A: Dilute 2.5 μg CAR (chimeric antigen receptor) DNA plasmid and 20 μL lentiviral packaging mix (ALSTEM, catalog number VP100; see Appendix B3) into 0.5 ml DMEM or Opti- 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 and 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. Add all the transfection...
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 necessary viral structural proteins, 3) expression of the vesicular stomatitis virus (VSV) carbohydrate Envelope carrier for protein (G). The third-generation lentiviral packaging system offers maximum biosafety because the lentiviral Rev gene is provided as an independent vector relative to other structural genes, further eliminating the possibility of reverse recombination of the vector into replication-competent viral particles. The third generation lentiviral packaging mix supports only chimeric 5' LTR lentiviral expression vectors in which the HIV promoter is replaced by CMV or RSV, thus making it TAT independent.
[0217...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More