mRNA-Lipid Nanoparticle Gene Delivery

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, Macrophages, 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 of 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 effects 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 CAR therapy. It typically uses electroporation for mRNA delivery, although it can be cytotoxic.

Lipid nanoparticles for mRNA delivery (LNPs) have been used in the pharmaceutical industry as a vehicle to deliver a variety of therapeutics. The mRNA delivery system was designed for ex vivo to human immune cell subtypes and many different formulations can be generated for the highest efficiency for different use cases. 

Our platform offers a completely validated mRNA delivery or mRNA LNP delivery system to many different cell types to induce functional protein expression and research the potential of mRNA and LNP-based gene therapies.

mRNA-LNP Gene Delivery Technology and Service Offerings

For more information, e.g. timelines and costs, speak with a technology specialist:

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Figure 1. Immune Cell Therapy Pipeline with mRNA-LNP. Overview of pre-clinical cell therapy development by using lipid nanoparticles for mRNA delivery, starting from antibody engineering, mRNA encapsulation, immune cell transfection & expansion, and its potential for clinical application.

Advantages of mRNA Delivery System LNP mRNA Delivery Applications
  • Transient expression system
  • Low immunogenicity
  • Easy to scale up and manufacture
  • Can carry large and complex payloads
  • Applicable to many avenues of therapeutic research
  • Vaccine development
  • Antibody delivery in-vivo
  • Chimeric antigen receptor (CAR) T/NK generation
  • Protein replacement therapies
  • Immune cell engineering: Macrophages, T cell, NK cell, B cell, Dendritic cell, Gamma Delta T cell


Service Plans and Prices 

Phase I mRNA design and synthesis by IVT $2,000
Phase II  mRNA production and purification. Validation by electroporation. $2,500
Phase III mRNA lipid nanoparticle formulation and encapsulation, followed by in vitro validation. $5,000
Optional Services mRNA-LNP scale up (up to 100mL)
mRNA-LNP validation in vivo
Antibody generation by LNP
Immune cell engineering by LNP
Please Inquire.


Effective Transient Gene Expression

Using the mRNA-LNP platform to deliver transient gene expression in many different cell types (HEK293, T cell, NK cell, dendritic cell, macrophage, stem cells, etc). The transient gene expression allows short-term gene expression delivered to targeted sites and is removed by the body after the delivery mechanism accomplished its tasks, for example:  
a.    mRNA-encoded non-mutated tumor antigens to prime the host immune system to defend against tumor cells.  
b.    Candidate mRNA-encoded cytokines to provide blueprint plans to the patient’s body to produce immune-activating/modulating cytokines that support anti-cancer immune response.  


Figure 2. K562 Transfected with GFP-LNP. Fluorescent imaging of K562 cells, before and after transfection with GFP+ mRNA-LNP.


Figure 3. HEK293S Transfected with GFP-LNP and Stability Test. 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

mRNA-LNP Immune Cell Engineering


Figure 4. GFP+ mRNA-LNP Delivery and Expression in T Cells. Left: High expression of GFP measured by flow cytometry 72 hours after adding GFP+ mRNA-LNP (PM-LNP-0021). Right: High cell viability measured by trypan blue cell counting.


Figure 5. CAR+ mRNA-LNP Delivery and Expression in NK Cells. 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.


Figure 6. CAR+ mRNA-LNP Delivery and Expression in T Cells. 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.

CAR-T Cells Generated with BCMA CAR mRNA-LNP 

Chimeric Antigen Receptor T cell and NK cell engineering can be easily generated with mRNA-LNP. CD19, BCMA, CD37, bispecific CARs, and other target tumor antigen CAR-T/NKs can be used for CAR validation or use in successfully treating relapsed cancer patients.

10 Components of the CAR-T/NK Platform

Figure 7. CAR-T cells were generated with BCMA CAR mRNA-LNP (PM-LNP-0017) 24 hours after LNP addition. 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: