ProMab’s mRNA-Lipid Nanoparticle (LNP) Gene Delivery Custom Service Platform
ProMab Biotechnologies, Inc., offers a comprehensive platform to advance immune cell therapy engineering for research applications. One of the most important questions that are asked in the field is the best method to introduce a novel gene into the body's natural defense force, our immune system. When we consider adoptive cell therapies we are including T cells, NK cells, Macrophage cells, Dendritic cells, and the other prevalent immune cell subtypes. Engineering them and delivering your gene of interest whether it be a chimeric antigen receptor (CAR) or a different transgene can be tricky and ProMab offers the solutions to assist researchers with many gene delivery options.
Chimeric antigen receptor (CAR) cell therapy relies on manipulating a patient's T cells ex vivo and creating highly effective cancer-targeting machinery that is shown to be capable in remission in patients with acute lymphoblastic leukemia and large B cell lymphoma. These methods have been widely used with viral delivery vectors which are longer lasting (generally permanent) as the virus used will integrate into the host cell's genome. This method could lead to severe adverse affects and thus other options for gene delivery are explored.
Messenger RNA (mRNA) has prospered as a strategy to generate transient gene expression in immune cells to mitigate the downsides associated with viral vectors, including for CAR therapy. It typically uses electroporation for mRNA delivery, although which can be cytotoxic.
Lipid nanoparticles (LNPs) were designed for mRNA delivery ex vivo to human immune cell subtypes and many different formulations were generated for the highest efficiency. Our platform has been tried and true to deliver mRNA to many different cell types to induce functional protein expression and indicate the potential of mRNA and LNP based gene therapies.
10 Components of the CAR-T/NK Platform
Immune Cell Therapy with mRNA-LNP
Figure 1. Overview of pre-clinical cell therapy development with mRNA-LNP technology, starting from antibody engineering, mRNA encapsulation, immune cell transfection & expansion, and it's potential for clinical application.
We can validate the positive cells with many different functional assays
K562 Transfected with GFP-LNP
Figure 2. Fluorescent imaging of K562 cells, before and after transfection with GFP+ mRNA-LNP.
HEK293S Transfected with GFP-LNP and Stability Test
Figure 3. Expression of GFP-LNP in HEK293S cells and their stability. Freeze-thaw cycles of HEK293S cells transfected with ProMab mRNA delivered via ProMab lipid nanoparticles. Flow cytometry measurement after 1 thawing cycle, and 24 hours at -80°C
We can deliver mRNA-LNP to immune cells
GFP+ mRNA-LNP Delivery and Expression in T Cells
Figure 4. Left: High expression of GFP measured by flow cytometry 72 hours after adding GFP+ mRNA-LNP. Right: High cell viability measured by trypan blue cell counting.
CAR+ mRNA-LNP Delivery and Expression in NK Cells
Figure 5. ProMab mRNA-LNP Nanoparticle with chimeric antigen receptor (CAR) mRNA generated CAR-NK cells. Left: Non-transfected NK cell control. Right: NK cell + LNP-CAR formulation.
CAR+ mRNA-LNP Delivery and Expression in T Cells
Figure 6. ProMab mRNA-LNP Nanoparticle with chimeric antigen receptor (CAR) mRNA generated CAR-T cells. Left: Non-transfected T cell control. Middle: T cell + LNP-CAR formulation by mouse Fab-APC. Right: T cell + LNP-CAR formulation, detected by mouse Fab-FITC.
ProMab BCMA-CAR T cells kill target cells in 24 hours after RNA CAR-LNP introduction
Figure 7. Left: BCMA CAR-T cytotoxicity measured by real-time cytotoxicity assay (RTCA). Right: Cytokine secretion measurement by IFN-gamma ELISA.
We also have off the shelf solutions that will quickly establish and advance your projects in the following areas: