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Fast and semi-automatic optimization of capture step for rAAV purification using CIM® SO3 96-well monolithic plate

First chromatographic step in industrial platform using monoliths is a capture step, where rAAV is selectively bound to strong cation exchanger (SO3) in acidic pH conditions (1,2). In this step majority of host cell contaminants are removed and rAAV is strongly bound to the matrix. Productivity of the step is therefore strongly influenced by the dynamic binding capacity (DBC). DBC is dependent on the sample preparation prior to chromatography and the availability of the binding sites at the chromatographic matrix.

Due to increasing demand for rAAV quantity and quality for clinical manufacturing, large volumes of upstream produced material are being purified on industrial scale. At this point, rAAV capture is a first chromatographic step to be optimized for optimal selection of the chromatographic parameters. Doing this, SO3 monolithic columns can be used with their full potential. To reduce the development time using multi-factor screening and increase comparability during downstream process development, CIM® SO3 monolithic 96-well plates were introduced to Sartorius BIA Separations portfolio. Obtained results on small scale can be applied to the CIMmultus™ line, which is scalable to large industrial volume.