The development of CAR T-cells is a complex process that requires multiple components in the workflow including I) screening and sequencing of mAbs that are specific to the cancer antigens; II) engineering and validation of ScFv and CAR of different varieties for their specificities and activities; III) production of high titer lentivirus for CAR constructs; IV) isolation, activation and expansion of primary T cells from specific donor or patient that exhibit a specific cellular phenotype; V) transduction of activated T cells with CAR-encoding lentivirus; VI) validation of engineered CAR T-cells through FACS and functional analysis.

BPS Bioscience has developed a growing list of CAR-T products including CAR-T lentiviruses, CAR-T reporter cell lines, and fully validated functional CAR T-cells as well as antigen-specific cancer target cells labeled with Firefly luciferase. In addition, we also have developed ELISA assays (kits) for cytokine (IFNγ) detection and Firefly luciferase-based cytotoxic assays for evaluating target-specific CAR-T activation and cancer cell killing.

BPS Bioscience is now offering custom CAR-T development services addressing multiple components of the CAR-T workflow mentioned above. This includes designing and constructing lentivector for scFv-CAR through validation of functional CAR T-cells of different varieties, including mono-specific CAR, dual CAR and tandem CAR. The services can be delivered in multiple milestones, as follows:

1. CAR Lentivirus Production and Initial Validation

• Design and construct lentivector for scFv-CAR of different varieties including mono-scFv CAR, dual scFv CAR and tandem scFv CAR

• Transfect lentivector and packaging plasmids in 293T cells to make high titer concentrated lentivirus

• Validation of the functional activity of CAR construct by transducing the CAR lentivirus in NFAT reporter Jurkat cell line to check for functional expression of CAR by FACS and reporter assay

2. T-Cell Preparation for CAR Lentivirus Transduction

• Isolate, activate and expａnd primary T cells (from healthy donors or patients)

• Transduce T cells and check the expression of CAR by FACS (CAR T-cells establishment)

3. Validation of Functional Activity of CAR T-Cells in Co-culture Assays

• Check IFNγ production in CAR T-cells using BPS Bioscience’s IFNγ ELISA kit

• Establish labeled target cells: cancer marker and luciferase double stable CHO cell lines

• Cytotoxic (cell killing) assay using co-culture of target cells (double stable CHO cell line) and effector cells (CAR T-cells)

• CAR T-cell expansion and optimization

BPS Bioscience's CAR T-cell development services can be applied to any of the following areas:

• Compound and Ab screening using CAR T-cells or CAR/NFAT reporter Jurkat cells

• Intracellular co-stimulatory and activation domain comparison

• Study CAR-T signaling in the tumor microenvironment

• Screening CAR-T donor variations

• Evaluate Treg and T memory cells in CAR-T setting

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