Sartorius BIA Separations
CIMmultus® OH chromatographic column relies on hydrophobic interaction chromatography (HIC) employing uncharged, OH ligand to provide an excellent solution for the purification of various large biomolecules, such as LNPs, virus particles, VLPs and extracellular vesicles. This column is uniquely suitable for large biomolecule purification due to the low shear stress imparted of the purely laminar flow.
Large biomolecules are retained in the presence of precipitating salts or polyethylene glycol at neutral pH and eluted with high resolution in order of increasing size with descending salt or polyethylene glycol. Choosing 6 µm channels is recommended to ensure low backpressure during the process.
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FAQ About CIMmultus® OH
Yes. We have developed a platform process for purifying bacteriophages on monoliths. The first, capture step is on OH column. It is a universal step tried on more than 20 different phages. The step concentrates phages, and removes host cell proteins, DNA in the presence of precipitating salts or PEG. For cruder samples or capture step we recommend 6 µm channel size. For polishing step, we employ either QA or PrimaS column chemistries.
Hydrophobic capture on OH column may not work with samples containing detergent (e.g. from lysis). This capture method is most suited for an excreted AAV (supernatant). It may also be evaluated for other types of samples.
Our recommended column for AAV capture is SO3.
Yes. CIMmultus OH chromatography column is an efficient alternative to traditional methods. It ensures rapid and robust LNP purification with very high recovery. With an uncharged ligand and broad pH range, it efficiently removes impurities, resulting in purified LNPs with higher size uniformity and improved mRNA quality.
Our primary recommendations for VLP testing are QA, SO3 and OH columns, depending on the pI and sample properties. In some cases, 2 chromatographic steps are needed; we recommend testing CIMmultus OH column first, following by ion-exchange column (typically QA or SO3 work best). Choosing the suitable chemistry depends on sample properties, stability and proteins, which make up the VLP. Based on our recent experiences, we have observed excellent results using OH as the capture step.
For cruder, viscous samples or capture step we recommend choosing the 6 µm channel size.
Yes, monolithic columns are designed for multiple uses, their lifetime depends on the sample and column maintenance. It is recommended to clean the column (at least 2h in cleaning solution) after every purification run. We recommend following the instructions written in the Instructions for Use and optimize the cleaning for your sample.
