Inside of mRNA therapeutics manufacturing facility

Development and commercialization of therapeutics involving new and emerging technologies present the market with new challenges, from establishing secure supply chains of critical raw materials to creation of customized solutions. At Thermo Fisher Scientific, we rise to this challenge by applying our breadth and depth of technologies and resources to support your efforts. Let us help select the right products for your applications.

Our solution to address all aspects of mRNA therapeutic development

Products

  • Comprehensive selection of Thermo Scientific TheraPure in vitro transcription reagents
  • Verified Invitrogen Dynabeads magnetic beads for superior analytical reproducibility
  • Extensive library of proprietary delivery reagents
  • Custom gene synthesis with GeneArt GeneOptimizer to maximize expression of your synthetic genes
  • Innovative POROS Oligo (dT)25 affinity resin for selectivity and capacity requirements

Capabilities

  • Scalable systems support early-stage research through clinical development and commercialization
  • Leading innovation driven by continual process optimization
  • Dedicated team of technical experts
  • Superior quality management system
  • Flexible licensing options
Synthesis

mRNA synthesis

  • TheraPure enzymes (animal origin-free and ampicillin-free)
  • NTPs and modified nucleotides
Purification
Formulation and delivery
  • Invivofectamine Rx Reagent

Accelerated development of mRNA-based therapeutics has necessitated the transformation of standard, research-grade in vitro transcription (IVT) reagents into raw materials specifically designed and manufactured under advanced quality standards (Table 1). TheraPure reagents for mRNA therapeutics provide our partners access to the most experienced development, analytical, and manufacturing teams in the industry.

Table 1. In vitro RNA synthesis solution.

ProductsQuality controlManufacturing

IVT reagents:

  • TheraPure T7 RNA polymerase
  • TheraPure RNase inhibitor 
  • TheraPure inorganic pyrophosphatase
  • ARCA (anti-reverse cap analog)
  • NTPs and modified nucleotides

Posttranscriptional modification:

  • TheraPure capping enzymes
  • TheraPure 2’-O-methytransferase
  • TheraPure Poly(A) polymerase

Template degradation:

  • TheraPure DNase I
  • ISO 13485- and ISO 9001-certified manufacturing facilities
  • Segment-tailored QC tests
    • Bioburden testing (microbiological contamination)
    • Endotoxin
    • Ribonucleases
    • Residual host DNA
    • Endo- and exo-deoxyribonucleases
    • Functional testing
    • Protein purity
  • Ability to scale manufacturing
  • Class 100,000 cleanroom standards (class D)
  • Custom formulation and testing
  • Custom development for nucleic acid-based molecules, including modified nucleotides and capping molecules
  • Custom filling, packaging, and labeling

Animal origin-free (AOF) and ampicillin-free (AF) enzymes

The TheraPure enzymes for in vitro transcription are manufactured under stringent animal-free and ampicillin-free processes. These enzymes are grown in media and purified completely without animal-sourced components. Our AOF manufacturing process eliminates potential prion (bovine spongiform encephalopathy and transmissible spongiform encephalopathy) and virus-derived disease risks. Components of animal origin are replaced with AOF components throughout the manufacturing process—from fermentation to enzyme formulation.

The absence of animal-derived materials in the manufacturing process minimizes the need for excessive traceability documentation and accountability. In addition, the AOF enzymes help to reduce the risk of ampicillin-related allergic reactions and other side effects.

Generation of mRNA by in vitro transcription includes several cleanup steps. Traditional purification techniques, like phenol:chloroform followed by precipitation or spin-column, have limited capacity with multiple steps and are not easily automated. On the other hand, magnetic bead-based technologies are flexible, scalable, and automation friendly.  An mRNA purification protocol based on Invitrogen Dynabeads MyOne Carboxylic acid and a proprietary binding buffer can deliver higher yield than traditional purification methods.

Purification by generic capture

  • Invitrogen Dynabeads MyOne Carboxylic Acid for generic capture of mRNA (Figure 1)
  • In-process purification
  • Optimized buffers available under confidential disclosure agreement
  • Increased mRNA concentration
Illustration depicting purification protocol of in vitro transcribed RNA samples in workflow steps of binding, washing, and recovery of mRNA

Figure 1. Capture workflow with Invitrogen Dynabeads MyOne Carboxylic Acid.

Binding capacity

  • Up to 1.6 mg mRNA per mg Dynabeads MyOne Carboxylic Acid
  • Flexible and scalable protocol with a high recovery rate (Figure 2)
Histogram showing how mRNA purification protocol based on Dynabeads MyOne Carboxylic acid can be scaled proportionally to desired reaction volumes with yield of 1 mg of mRNA per mg of beads, unlike spin column which is not scalable

Figure 2. High mRNA recovery rate as determined using the Invitrogen Qubit 4 Fluorometer.

As a world leader in providing in vitro transfection reagents, our committed team of scientists continually develops evolving technologies to meet the growing demands for the latest mRNA delivery formulations, especially for therapeutic applications. Our in vivo mRNA delivery platform provides optimal organ delivery with minimal toxicity in small animal models (Table 2).

Table 2. In vivo mRNA delivery solution.

TechnologyInvivofectamine Rx mRNA
PotencyIn vivo luminescence (1 x 1010 photons/sec) at 1 mg/kg mRNA dose
ToxicityLow toxicity
Specificity*Liver, spleen, lung, intramuscular—other tissues may be accessible*
ScalableYes
GMP*Under development*

* Please inquire via email

Optimized proprietary delivery

We utilize a combinatorial method to identify novel compounds and optimize formulations specific for different RNA payloads through proven design of experiment (DoE) and extensive proprietary know-how in the field of transfection (Figure 3).

  • Extensive screening of potential candidates to select the optimal LNP formulations
  • DoE for optimizing LNP formulation process parameters 
Illustration of design of experiments to screen, identify, and optimize lipid nanoparticles formulations for in vivo RNA delivery platforms

Figure 3. Schematic of design of experiment (DoE) and formulation screening.

High-efficiency mRNA delivery

Invitrogen Invivofectamine Rx Reagent provides a method for in vivo mRNA delivery (Figure 4), allowing our partners to accelerate their research and development of new technologies.

  • Multicomponent, proprietary lipid nanoparticles
  • Uniformly sized (90 nm) >90% encapsulation efficiency
  • Potency demonstrated by in vivo luminescence (1 x 1010 photons/sec) at 1 mg/kg mRNA dose (Figure 4)
  • Specificity shown in liver, spleen, lung, and muscle
  • Scalable process
Panel A is fluorescence imaging showing sequential performance improvement of Invivofectamine Rx reagent for in vivo delivery of mRNA in mice livers  Panel B is a histogram quantifying the fold change observed by fluorescence imaging post-IV injection into mice

Figure 4. Sequential improvement in performance of reagents through DoE. in vivo delivery of luciferase mRNA (FLuc) using Invitrogen Rx mRNA demonstrating continuous improvement in reagent delivery performance as shown in (A) visual and (B) quantitative assessment of luciferase expression 4 hours post-IV injection in mice.

Related areas

mRNA purification solution   |   Gene therapy research   |   OEM and commercial supply   |   In vitro transcription

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