mRNA Drug Production

Messenger RNA (mRNA) acts as an essential intermediary between the information stored in DNA and protein synthesis. It is a single-stranded ribonucleic acid (RNA) molecule that is created in the cell nucleus to match the sequence of genetic information in DNA. mRNA then moves into the cytoplasm, where protein synthesis occurs. mRNA is often compared to a recipe or a blueprint in that it contains the directions for making a specific protein.

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mRNA therapies, including vaccines like those deployed against COVID-19, are valuable tools with a growing list of potential applications in modern medicine. They carry mRNA strands, instructing cells to make a particular protein. This target protein can act to treat or prevent disease. The purpose might be to protect against a virus, like the flu or COVID- 19, or to attack cancer cells or treat a rare genetic disease by replacing a defective protein. mRNA vaccines and other mRNA drugs have shown great promise, and their market is expected to expand rapidly in the coming years. Globally, the mRNA therapeutics market size was valued at $39.90 billion in 2021 and is expected to grow at a compound annual growth rate of 1.7% from 2022 to 2030.

The following qualities make mRNA drugs and vaccines attractive options:

• Safety: There is no risk of the drug integrating into the genome or causing infection. It is naturally degraded in the body.
• Efficacy: The widespread use of mRNA vaccines against SARS-CoV-2 demonstrated the superior efficacy of this technology compared to other vaccine platforms.
• Development Speed: Compared to the traditional vaccine development timeline of 10 to 15 years, mRNA vaccines offer a much shorter and simpler development process, from design to manufacture.
• Broad Potential: mRNA drugs can potentially treat a wide range of diseases. Due to the continuous translation of the mRNA code, this approach can spur sustained protein expression compared to other drugs.
• Scalability: The bioprocesses to manufacture mRNA therapies are easily scalable.

Vaccines account for most of the mRNA therapeutic products in development, but the biomedical uses for mRNA-based medicines are rapidly evolving. In addition to vaccines, there is potential to develop a wide range of mRNA therapies, including mRNA-based cell and gene therapy, cancer vaccines and immunotherapy, and protein replacement therapies. Infectious diseases that could be mRNA vaccine targets include influenza, respiratory syncytial virus, Zika, rabies, and Ebola.

The manufacture of mRNA drugs involves a series of production steps. These include:

• Plasmid DNA Production and Linearization: Production of mRNA therapies begins with a plasmid DNA (pDNA) template. pDNA is a small, circular DNA fragment containing the genetic blueprint for the desired protein. This step involves bacterial fermentation and subsequent purification to isolate the pDNA, which is then linearized for the next step.
• Purification: Chromatography is used to purify the linearized pDNA.
• In Vitro Transcription (IVT): A linearized pDNA is used as a DNA template and transcribed into mRNA. This is achieved through a process called in vitro transcription using RNA polymerase and other enzymes. This step requires precise temperature and pH control.
• mRNA Purification: The synthesized mRNA must be extracted from other molecules and impurities. Tangential flow filtration (TFF) is an efficient mRNA separation technique. Chromatography methods can be used as an alternative to TFF and for additional purification.
• mRNA Modification: Following transcription, the linear mRNA strand must be modified or “capped.” An enzymatic reaction replaces the “free” end of the RNA transcript with a molecular cap. The cap is essential to the cell’s ability to recognize and translate the mRNA into a protein. It also protects the mRNA from degradation.
• Lipid Nanoparticle Encapsulation: The purified mRNA is then formulated into a suitable delivery system. Lipid nanoparticles (LNPs) or other carriers create a protective coat for the fragile mRNA and deliver it to cells. For mRNA LNP, lipids dissolved in ethanol are combined with mRNA dissolved in an aqueous buffer in a Jet mixer, T-mixer, or Y mixer.
• Clarification Filtration and Sterile Filtration: Multiple steps are necessary to ensure the purification and safety of the final product.

When manufacturing mRNA drugs, it is essential to maintain mRNA stability throughout the process, ensure efficient encapsulation into LNPs, and avoid contamination from extractables and leachables (E&L). These hurdles can be effectively addressed with the right equipment.

Peristaltic pumps offer low-shear, accurate, reliable fluid handling, which is critical in mRNA production as drug products must be securely transferred between production steps. Nearly 80% of mRNA manufacturing processing steps use peristaltic pumps and require high-performance tubing. The value of the drug product increases as the process continues, as does the cost of product loss. By the formulation step, the value of a lost batch could exceed one million dollars. It is essential that the pumps and specialized peristaltic pump tubing used in these processes be designed to meet the stringent demands of mRNA production.

Unlike conventional extruded pump tubing, which may balloon or rupture at elevated pressure, GORE STA-PURE Pump Tubing, Series PCS employs a unique, patented composite structure that overcomes this limitation. It enables flow stability at higher operating pressures for tangential flow filtration (TFF) and chromatography steps with low shear peristaltic pumps. Reduced risk of pre-mature tube failure translates to maximized productivity for mRNA purification. Series PCS high pressure pump tubing (up to 100 psig) offers:

Conventional pump tubing may have limited resistance to aggressive organic solvents used in LNP encapsulation. But the patented composite of GORE STA-PURE Pump Tubing, Series PFL, is engineered to resist harsh chemicals, including organic solvents. Series PFL tubing enables flow rate stability up to 60 psig with a low extractables profile for the LNP encapsulation step, thereby contributing to maximized quality for mRNA drug delivery systems. Series PFL offers:

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