Messenger RNA

The most comprehensive chromatographic toolbox for purification of mRNA - from affinity to reverse phase.

CIM® technology offers a uniquely capable purification toolbox for rapid process development and scale-up to ensure:

  • The product has a intact poly(A) tail.
  • dsRNA and DNA template are removed.
  • Process and product-related impurities, such as enzymes, DNA and RNA fragments are removed.

Messenger RNA is large for its mass. It is also sensitive to shear forces. The large channel structure of the chromatographic monolith allows free access for large particles, and laminar flow prevents shear damage. The structure enables purification of mRNA up to 10 kb or larger.

Flow through monolith and porous particle

The rich spectrum of chemical properties of mRNA is the basis for a diverse selection of purification columns.  Messenger RNA contains a charged phosphate backbone, and hydrophobic nucleotide residues with hydrogen-bonding ability. 

Structure of RNA

Charge interactions, hydrogen bonding and other molecular forces also impose challenges. Messenger RNA acts as nucleation center and readily forms associations with other components in IVT mixtures. These heteroaggregates depress capacity, recovery, and the purification ability of every known purification method, including filtration, precipitation, and chromatography.

Toolbox overview

BIA Separations offers a comprehensive toolbox of chromatography columns to help you achieve the desired product quality, with the speed and ease of use provided by the monolith.

BIA Separations mRNA toolbox

SDVB and dsX Beta are avaialable for evaluation. For more information, contact us.

Product details

CIMmultus™ Oligo dT provides a single-step affinity capture of mRNA. Binding occurs through hydrogen bonding, with the oligo dT forming a stable hybrid association with the terminal poly(A) sequence of the target. Non-RNA contaminants flow though the column during sample loading.

Oligo dT purification of mRNA

CIMmultus™ C4 HLD is a hydrophobic interaction chromatography monolith for purification of nucleic acids. Sample is applied at high ionic strength in salts that precipitate RNA. The majority of DNA, dsRNA and short transcripts elute earlier than intact ssRNA. Proteins and aggregates are typically eluted eluted by a NaOH cleaning step.

C4 HLD purification of mRNA

SDVB and dsX Beta are avaialable for evaluation. The content below provides a snapshot into their capabilities. For more information, contact us.

SDVB is a styrenedivinylbenzene-based reverse phase chromatography monolith for purification of ssRNA. DNA, dsRNA, and ssRNA are fractionated according to length. Selectvity for separation of double stranded and single-stranded species of similar length is influenced by the degree of base pairing.

SDVB elution

dsX Beta purifies large single-stranded mRNA (ssRNA) under aqueous conditions at ambient temperature. It can be used for polishing or as high resolution capture method. It removes dsRNA, DNA, proteins, and endotoxins while fractionating ssRNA in order of increasing size.

dsX elution chromatogram

SDVB and dsX Beta are avaialable for evaluation. For more information, contact us.

Custom ligand immobilisation

If our standard products are not within your required specification, other ligands can be coupled to the monolith. Our R&D can immobilise custom oligo sequences, proteins or enzymes. You provide the ligand, we immobilise it. Your process may benefit from:

Immobilisation of custom oligo sequence 

for affinity purification of RNA. Rapid mass transfer in the monolith allows affinity capture at short residence times. This enables efficient chromatography purification of large biomolecules. Custom and priprietary oligonucleotide sequences may improve the purity of the product, and can be immobilised on the monolith matrix.

See our application note for more information:

Immobilisation of enzyme 

for DNA/RNA or protein digestion, or plasmid linearisation. Immobilised enzymes can reduce enzyme use and cost, eliminate an ezyme removal step, and improve the digestion efficiency.

The following literature will provide more information on enzyme immobilisation:

With our customer-oriented approach we will ensure that the final product meets the needs for your development and process requirements. Get in touch through our contact form.