Allergy Tolarization With mRNA Therapeutics

Equipped with a team of professional scientists, Seattle Genova is capable of providing 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. With our comprehensive experience and advanced platform, we can give a series of mRNA-based replacement therapy services. We are excited to partner with our clients on the journey of bringing novel mRNA-based therapies to clinical application.

mRNA vaccines open the area for a safety-optimized prophylactic vaccination against allergic disorders. Future studies relating long-term effects and vaccine-induced versus natural immune responses will be desired to transfer this information to the clinics.

Conventional, as well as self-replicating mRNA vaccines, have demonstrated their potential to avoid the induction of an allergic phenotype in terms of allergen-specific IgE, allergy-associated cytokine profiles, eosinophilic lung infiltration, and airway hyperreactivity. Preliminary data boost the question of whether TH1 immune deviation induced by mRNA vaccination resembles the natural phenotype of a specific proportion of nonatopic individuals. Reservations regarding Good Manufacturing Practices manufacture expenses, shelf life stability, and absence of immunogenicity due to rapid in-vivo degradation have been surviving by novel findings.

Allergen‐specific immunotherapy, which is conducted by subcutaneous injection or sublingual application of allergen extracts, illustrates a helpful treatment against type I allergic diseases. However, due to the long period and adverse reactions, only a minority of patients agree to suffer this treatment. Alternatively, early prophylactic intervention in young children has been formulated to stop the rise in patient numbers. Plasmid DNA and mRNA vaccines encoding allergens have been indicated to induce T helper 1 as well as T regulatory responses, which modulate or counteract allergic T helper 2–biased reactions. With respect to prophylactic immunization, additional safety measurements are needed.

In contrast to crude extracts, genetic vaccines give the allergen at great purity. Also, by targeting the encoded allergen to subcellular compartments for degradation, the discharge of native allergen can be avoided. Due to inherent safety characteristics, mRNA vaccines could be the candidate of option for preventive allergy immunizations. The subtle priming of T helper 1 immunity induced by this vaccine type nearly resembles responses of non‐allergic individuals and—by increasing via natural allergen exposure—could suffice for long‐term safety from type I allergy.

STEP-BY-STEP PROCESS 

Step 1. Plasmid Manufacturing 

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

Step 2. Transcription 

In-vitro transcription (IVT)

In vitro transcription is a method that facilitates the 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 utilizing the corresponding RNA polymerase.

Step 3. mRNA purification

After certain manufacturing steps it is significant to purify the mRNA. 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.

Step 4. 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.

Step 5. QC release and stability testing

Relying on your final mRNA application and clinical stage, the quality control testing requirements may vary.

◆ 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 assistance at competitive prices

◆ Custom tailored assistance to meet specific application or program needs

◆ Vast variety of modification, treatment, and purification options