Price/availability/specifications subject to change without notice. Unless otherwise indicated, our catalog and customized products are for research use only and not intended for human or animal diagnostic or therapeutic use.
Or leave a message with a formal purchase order (PO) Or credit card.
Chimeric Antigen Receptor T-Cell Service
DNA vectors used in the CAR T-cell generation service will NOT be transferred to clients under this arrangement.
Custom Chimeric Antigen Receptor (CAR) T-cells are generated by experienced scientists working on antigen and antibody production, antibody engineering, lentivirus, and cell line development services for decades. T cell receptors are engineered to graft an arbitrary specificity, such as that of a monoclonal antibody, onto an immune effector cell, such as T cell. If T cells are removed from a patient and modified to express receptors specific to the particular form of cancer, the engineered T cells recognize and kill the cancer cells after being reintroduced into the patient. The strategy has been shown to be somehow effectivie in hematologic malignancies.
Generation of CAR T-cells:
The most common form of chimeric antigen receptors (CARs) are fusions of single-chain variable fragments (scFv) derived from monoclonal antibodies (also called ectodomain) to CD3-zeta transmembrane and endodomain. The general process of CAR T-cell generation includes:
- Antigen (peptide or protein) production;
- Mouse, rat, or hamster monoclonal antibody production;
- Sequencing hybrioma antibody;
- Chimeric scFv production and validation;
- Antibody humanization;
- Molecular construction of CAR cassette, lentivirus packaging and production;
- T-cell activation, transduction, and CAR T-cell expansion;
- Confirmation of CAR expression, IFN-gamma ELISA, and cytotoxicity assay of transduced and expanded T-cells.
Antibody humanization is not needed if a murine-specific scFv is used with immunodeficient mouse tumor models for proof-of-concept studies.
Fully human antibodies can be obtained by screening phage or yeast libraries or sequencing human repertoire. Then the process from murine monoclonal antibody development to antibody humanization can be replaced except chimeric scFv production and validation.
Camelid single domain antibodies (VHH) or nanobodies can be used to replace scFv.
In the endodomain of the first generation of CARs, there is only the intracellular domain from the CD3 zeta-chain, the primary transmitter of signals from endogenous TCRs. In the second- and third-generations of CARs, intracellular signaling domains from various costimulatory protein receptors, such as CD28, 41BB, ICOS, and OX40, are added indivivually or in combination to provide additional signals to the T cell and to improve the antitumor activity of T cells.
Service of molecular construction of lentivirus CAR cassette and CAR T-Cell generation includes:
a. Molecular construction of CAR cassette, lentivirus packaging and production;
b. T-cell activation, transduction, and CAR T-cell expansion;
c. Confirmation of CAR expression, IFN-gamma ELISA, and cytotoxicity assay of transduced and expanded T-cells.
a. 2 vials of the expanded T-cells (5 x 10^7 cells per vial, a mixture of CAR T-cells and non-transduced T-cells);
b. CAR T-cell generation study report including CAR design and confirmation of CAR expression in CAR T-cells.
Fast turnaround: 10-12 weeks.
Syd Labs also provides the following viral-based cell line generation services:
Lentiviral Based Stable Cell Line Generation Services: development of stable cell lines using lentivirus.
Knock-out and Knock-in Cell Line Generation with CRISPR Genome Editing: development of stable cell lines for precise genomic modification.
Syd Labs also provides the following stable cell line generation services:
Standard Stable Cell Line Development Services: non-viral generation of adhension or suspension stable cell lines with one or two gene insertion.
Stable Cell Line Development Services for Protein Production: development of stable cell lines for recombinant protein expression.
Stable Cell Line Development Services for Antibody Production: development of stable cell lines for recombinant antibody expression.
Stable Cell Line Development Services for Biologics Production: generation of stable cell lines for manufacturing recombinant protein or antibody drugs.