Optimizing AAV Purification: Comparative Evaluation of Three CIMmultus^® Polishing Columns for Enhanced Vector Enrichment and Final Product Purity

As the demand for high-quality AAV products increases, manufacturers face the dual challenge of ensuring both purity and efficiency in their processes. To evaluate and improve the polishing step following the capture phase in the AAV purification process, three monolithic polishing columns, CIMmultusQA HR, PrimaT, and PrimaS, were evaluated for their performance. The QA column acts as a strong anion exchanger, efficiently targeting negatively chargedions. PrimaSis a multimodal ligand that combines hydrogen bonding with anion exchange chromatography for versatile separations. PrimaTfurther enhances this versatility by offering weak-anion exchange, hydrogen bonding, and metal affinity coordination, making itsuitable for complex molecular separations. While the QA HR column is already well-known in the biotechnology field, PrimaTand PrimaSare just beginning their journey. One of the key advantages of PrimaTis its ability to separate different subpopulations of full and empty capsids. PrimaS, on the other hand, distinguishes itself from the other two columns by utilizing a pH gradient, which can sometimes be the optimal choice for the polishing step.