mRNA-based Cell Therapy Services

Equipped with a team of professional scientists, Seattle Genova is capable of giving specialized support in the design, production, and evaluation of mRNA formulation services. Our mRNA manufacturing template gives a robust workflow, while our lipid nanoparticle (LNP) manufacturing processes ensure a high-quality and consistent supply. 

The therapeutic use of messenger RNA (mRNA) has fueled great hope to combat a broad range of incurable diseases. Recent rapid progress in biotechnology and molecular medicine have facilitated the generation of nearly any functional protein/peptide in the human body by introducing mRNA as a vaccine or therapeutic agent. This illustrates a rising precision medicine field with enormous promise for preventing and treating many intractable or genetic diseases.

CAR T cell therapy (or chimeric antigen receptor T cell therapy) is an illustration of cell-based gene therapy. This type of treatment integrates the technologies of gene therapy and cell therapy. Cell therapy introduces cells to the body that have a specific function to encourage treat a disease. In cell-based gene therapy, the cells have been genetically altered to provide them the special function. CAR T cell therapy instructs a gene to a person’s T cells, which are a type of immune cell. This gene provides instructions for making a protein, called the chimeric antigen receptor (CAR), that attaches to cancer cells. The modified immune cells can mainly attack cancer cells.

Applications

The CAR-T cell has been utilized predominantly in the treatment of hematological malignancies, including acute lymphoblastic leukemia, chronic lymphocytic leukemia, lymphoma, and numerous myeloma. Solid tumors containing melanoma, breast cancer and sarcoma offer enormous promise in CAR-T cell research and development. 

PRODUCTION PROCESS 

DNA plasmid production for mRNA synthesis

mRNA synthesis begins with plasmid design and production. Plasmids are generated in bacterial cultures, then harvested and purified.

In-vitro transcription (IVT)

In vitro transcription is a method that facilitates for template-directed synthesis of RNA molecules of any sequence from short oligonucleotides to those of various kilobases in μg to mg quantities. It is based on the engineering of a template that contains a bacteriophage promoter sequence (e.g. from the T7 coliphage) upstream of the sequence of interest fulfilled by transcription using the corresponding RNA polymerase.

mRNA purification

mRNA purification eliminates enzymes, remaining nucleotides, plasmid DNA, and defective mRNA. New emerging technologies like Fibro chromatography, currently accessible for mAb purification, are in development for molecules such as DNA plasmids and mRNA.

mRNA encapsulation and polishing

The purified mRNA-based therapeutic is formulated in lipid nanoparticles (LNPs) as a drug delivery vehicle. Core chromatography can be used to further eliminate impurities.

QC release and stability testing

RNA content by UV-Vis

Purity by IRRP HPLC

Residual DNA by RT-qPCR

Residual protein by MS

Potency by cell-free translation

Endotoxin and residuals measurements

Sequencing

SERVICE HIGHLIGHTS

◆ High quality products and services at competitive prices

◆ Custom tailored support to meet specific application or program needs

◆ Wide variety of modification, treatment, and purification options

◆ Affordable custom synthesis up to gram scales of mRNA and long RNA (multiple kilobases)

◆ In-house plasmid manufacturing optimized for therapeutic mRNA production

DELIVERABLES

We provide high throughput evaluations along with faster results. In addition to that, we provide many different mRNA formulation services to meet your various end-point in vaccine delivery. These formulations come with specific functionality to improve the efficiency of vaccines in the physiological environment.

REFERENCES

1.Mitchell, D. A. et al. Selective modification of antigen-specific T cells by RNA electroporation. Hum. Gene Ther. 19, 511–521 (2008).

2.Su, Z. et al. Telomerase mRNA-transfected dendritic cells stimulate antigen-specific CD8+ and CD4+ T cell responses in patients with metastatic prostate cancer. J. Immunol. 174, 3798–3807 (2005).

3.Holtkamp, S. et al. Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells. Blood 108, 4009–4017 (2006).