Nucleic Acid Purification Solutions

Improving the purification of mRNA and plasmid DNA

The increasing number of approved nucleic acid therapeutics drives the demand for efficient platform technologies to manufacture these new therapeutic modalities. We have built a toolbox of high-performing chromatography resins to help address your needs and help you increase efficiency and productivity in the purification of mRNA and plasmid DNA.

Simplify your mRNA purification process

One challenge we can help you overcome is obtaining larger quantities of synthetic mRNA to support clinical manufacturing. POROS™ Oligo (dT)25 Affinity Resin has been designed for the purification and isolation of mRNA from in vitro transcription (IVT) manufacturing processes. This affinity chromatography resin offers high selectivity and allows for easy mRNA purification. Through the AT-base pairing mechanism, the resin effectively separates mRNA from components of the IVT process, such as enzymes, unreacted nucleotides, partial transcripts, and plasmid DNA.

Graphical representation of POROS Oligo (dT)25 resin binding to a poly-A tailed mRNA molecule through AT base pairing

Figure 1. Mechanism of action of the POROS Oligo (dT)25 affinity resin. The Poly-dT ligand allows binding with poly-A tailed mRNA molecules through AT base pairing.

Why choose the Oligo (dT)25 affinity resin?

  • Easy mRNA purification from crude transcription mix
  • High dynamic binding capacity and high recovery
  • Ease of use – load in NaCl and elute in water or low salt buffers
  • Simplified workflow helps maximize efficiency and reduce the complexity of subsequent polish steps
  • Excellent scalability, enabling purification from benchtop to commercial manufacturing
  • Enables a single platform purification solution to all mRNA products holding a Poly-A tail

Chromatogram showing efficient separation of mRNA from an in vitro transcription mixture using POROS Oligo (dT)25 affinity resin

Figure 2. Chromatogram showing efficient separation of a 2000nt mRNA from an IVT mixture at a load concentration of 2 mg/mL. Elution was performed using H2O and yielded in >95% recovery.

Resins and reagents for the purification of mRNA – ordering information

Efficient and scalable purification of mRNA using affinity chromatography

Optimize your plasmid DNA purification process

The rapid growth of the gene therapy pipeline as well as genetic vaccination for various infectious diseases, requires larger scale production of plasmid DNA (pDNA) at high quality. POROS AEX resins are ideally suited for the purification of larger molecules such as plasmids and will help you overcome the common challenges of plasmid purification.

Plasmid purification challengePOROS resin features
Products and contaminants are similar in size50-micron bead size—providing superior resolution, tighter peaks and improved impurity clearance
Shear sensitivity and high viscosity limits operational flow ratesPoly(styrene-divinylbenzene) backbone — resulting in linear and scalable performance. The beads are rigid and have a high mechanical strength enabling easy and reproducible scale-up.
Conventional chromatography resins exhibit low binding capacities for pDNALarge throughpores — leading to a reduced mass transfer resistance compared to other resins. Capacity and resolution are maintained over a wide range of linear velocities, thereby establishing a more efficient purification process.
Plasmids are generally much larger than proteinsLarge throughpores — increased surface area accessible which favours the purification of larger molecules
POROS resins exhibit high capacity, high throughput, and superior resolution capabilities

Plasmid DNA dynamic binding capacity study

During a design of experiments (DoE) study, POROS AEX resins where tested using various process conditions to determine dynamic binding capacities for plasmid DNA binding. Download the article below to learn more about the complete study.

POROS ResinPlasmid DBC10% (mg/mL resin)
POROS D5015.6

Table 1. Dynamic Binding Capacity Determination using process conditions from DoE study. Process conditions: residence time 2.5 min, 1 mL column, 45 mS/cm conductivity

Optimizing downstream purification of high-quality plasmid DNA for gene therapy and vaccine production

A design of experiment (DoE) study was conducted in order to evaluate POROS AEX resins for pDNA capture, with the goals of optimizing process conditions to help maximize purity and recovery, determine the dynamic binding capacity (DBC) of POROS AEX resins for pDNA,and confirm optimal operating parameters.

Learn more

Article for optimizing strategies for pDNA purification

Plasmid Purification Resins

mRNA and pDNA polishing

For subsequent polishing steps after mRNA or plasmid DNA capture, we recommend our suite of differentiated novel Hydrophobic Interaction Chromatography resins.

Related resources

Bioprocessing resources

Access a targeted collection of scientific application notes, case studies, posters, white papers and more for bioprocessing:

Pharmaceutical Grade Reagent. For Manufacturing and Laboratory Use Only