CAR-T-Cell-Based-Immunotherapy-Development-of-CAR-T-Cells

In the CAR-T cell constructing CAR-T CELL-BASED IMMUNOTHERAPY processes, the first step is to eukaryotically express the DEVELOPMENT OF CAR-T CELLS target antigen. Mice are vacci- nated with the antigen, and 1 Flow Chat of CAR-T cells generation the B cells are collected to generate hybridomas with Antigen Expression Tumor Immunize B cells Hybridomas myeloma cells using cell gene vector antigen mouse fusion technology. After sever- al rounds of antigen-specific CAR vectors VH & VL DNA RNA Screening screening, hybridomas with the highest affinity with the +/– CD4&CD8 target antigen are collected, T cells and the total RNA is isolated Expansion IL-2, IL-7, Beads or and reverse transcribed for IL-15, IL-21 artificial APCs Flow sorting sequence analysis. Sequenc- Transduction column separation es of the antibody variable with CAR region are cloned into CAR vectors, including lentivirus, Pheresis Total white retrovirus, and transposome cells systems. Total white cells are Quality Return collected from the patient’s control to patient blood by pheresis methods CAR-T cells and separated with flow sorting to obtain the CD4 and 2 Vector Systems for CAR Engineering CD8 T cells, which are then in vitro matured and expanded. Lentivirus System After infection or transfection, the CARs are expressed in the 5’LTR Transfer plasmid of 1G and 2G 3’LTR Envelope plasmid of 1G and 2G T cells to finally generate the U3 R U5 Prom. VSV-G pA CAR-T cells. Ψ/RRE Prom. CAR U3 R U5 Lentivirus is HIV-1 based and Prom. Packaging plasmid of 1G rev nef pA the most mature system for T gag pol vif vpr tat cell engineering. The first (1G) vpu RRE and second (2G) generation lentivirus systems both consist Packaging plasmid of 2G rev of three parts: transfer, enve- tat lope, and packaging plasmids. Prom. gag pol RRE pA A regulator plasmid, which regulates viral transcription 5’LTR with no U3 Transfer plasmid of 3G ΔU3 3’LTR Envelope plasmid of 3G and nuclear export of Prom. R U5 Ψ/RRE Prom. CAR R U5 transcripts, is supplemented Prom. env pA to form the third generation. Gamma retrovirus system is Packaging plasmid of 3G Regulatory plasmid of 3G quite similar to lentivirus except for few elements Prom. gag pol RRE pA Prom. rev pA including promoters. Trans- posons are dual component Retrovirus System Transposome System systems composed of one plasmid carrying the CAR 5’LTR Transfer plasmid 3’LTR Donor plasmid (transposon) and the other U3 R U5 TIR Prom. CAR carrying the transposase. Ψ CAR U3 R U5 TIR Creative Biolabs Donor plasmid Envelope plasmid Transposase expression plasmid CAR/TCR-related gag pol Enve. Prom. Transposase Products & Services WHAT WE DO: Products: Diseases Associated Antigen One-Stop CAR-T Therapy Development Services CAR Vector System TCR Modified T Cell Development Services Viral Particle TCR-Like Antibody Services CAR/TCR Development Kits Dendritic Cell Vaccine Development Services Bispecific TCR Development Service Copyright © 2018 Creative Biolabs. All Rights Reserved. | Contact Us