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Implementing Chromatographic Methods for Evaluation of Large Scale Monolithic Columns for AAV Capsids Separation

Quaternary amine (QA) modified monolithic columns are frequently used for purification of therapeutically relevant 
adeno-associated virus (AAV) vectors, used in gene therapy. Beside process- and sample-derived variability, 
chromatographic material variability can influence the efficiency and scalability of AAV downstream processing. This study 
presents the evaluation of highly reproducible (HR) QA-modified CIM monolithic columns, where the goal is to elute AAV 
capsids in a very narrow conductivity range, regardless of the batch and size of the the column used. 
We have developed a chromatographic test method for proving the intra- and inter-batch homogeneity of the material 
through different column sizes. The method is based on separation of AAV2/8 capsids in ascending KCl gradient on 
CIMmultus QA 1 mL columns and specimen 0.2 mL units, taken from large-scale QA monoliths up to 8000 mL in size.