In nearly 5 years we have developed over 30 types of CAR-T/NK cells and target
cells with several patented innovations in the CAR construct design. We can
construct a CAR based on your antibody sequence or we can generate the entire
construct from a mouse monoclonal antibody or fully human single-chain
variable antibody fragment (scFv) from our existing library.
ProMab Biotechnologies Custom Chimeric Antigen Receptor Cell Engineering: A
complete service, from antigen to clinically relevant solution.
CAR-encoding lentivirus preparations are used to transduce human T cells.
Mononuclear cells are isolated from peripheral blood, then T cells are
activated with IL-2 and CD3/CD28 macrobeads (cat. # PM-CAR2002) and
transduced with lentivirus 24 hours and 48 hours following activation. Cells
are then passaged at 2-3 day intervals to maintain proper cell density.
Starting on day 8, CAR expression can be detected by flow cytometry, using an
anti-mouse or anti-human Fab antibody (or an anti-tag antibody if the CAR is
tagged). By day 12 there are enough cells to test the CAR in cytotoxicity and
cytokine-induction assays.
The process of generating and expanding CAR-T cells has been
improved with an optimized CAR-T expansion media (cat.#
PM-CAR2000).
We also screen human PBMC donors from the Stanford University
Blood Center. These donors can be used for screening of different
markers, checkpoint inhibitors, genomics, proteomics and metabolic
analyses. The donors can also be used for antibody and drug
discovery projects, to study drug resistance mechanisms and for
the discovery of novel biomarkers. These cells can be used to
develop personalized medicine or precision medicine approaches.
Our T cell activation and expansion platform mimics what occurs in
nature, but with greater efficiency and control.
Figure 1. Scheme of T cell activation in nature (top) and in ProMab's
platform (bottom)
Figure 2. Representative growth profile of transduced cells.
Figure 3. Morphology of transduced cells(day 10).
2~3x109 cells can be produced using ProMab's CAR-T medium (#PM-CAR2000) and
CD3/CD28 T cell activation beads (#PM-CAR2001-2003) (Figure 1). The growth of
the cells is monitored by counting (Figure 2) and microscopy (Figure 3).
CAR-T cells can subsequently be purified and isolated by ProMab
Biotechnologies CAR-T Extraction Kit.
Figure 4. Scheme of CAR-T cell isolation using ProMab Biotechnologies'
antibody-coated magnetic beads
We have also quantified that the increase of multiplicity of infection (MOI)
correlates to an increase in transduction efficiency and CAR expression.
Figure 5. Comparison by flow cytometry of tagged CAR-T cells generated at
either a MOI of 5 or 10.
Mouse monoclonal antibodies
Human monoclonal antibodies
Hybridoma Sequencing
Other Components of the CAR Platform:
