Method Optimization & High Throughput Purification for mRNA and rAAV

Speaker:
Marko Narobe, Sartorius BIA Separations

Abstract:
The downstream processing of virus particles, vesicles, RNAs, plasmids and other forms of DNA, contains multiple interdependent steps, each requiring optimization for best results.  This webinar will showcase how to shorten the development time by screening multiple conditions at once, with small sample intake and process automatization. In these processes we used our newly launched CIM monolithic plates.

The CIM monolithic plates are a standard plate design, manufactured according to ANSI standard. In each well there is a defined amount of monolithic chromatographic media. Due to its intrinsic properties the mass flow through the monolith is convective. This enables us to have fast processes, and no shaking or incubation is required with our plates.

First, we present the screening of different mobile phases for rAAV capture step. Optimization of capture step leads to increased process productivity and product purity, as well as improves the polishing step. High vector recovery and greater reduction of impurities translate to preparative scale.

Subsequent part of the webiar focuses on the importance of finding conditions that increase dynamic binding capacity (DBC) of mRNA on Oligo dT. Screening experiments help to to identify the factors that affect the binding capacity and achieve a DBC of >6 mg/mL.

In both cases, the optimized conditions were scaled-up to preparative scale chromatography, resulting in successful implementation of screening tools for process development optimization. Watch the webinar to learn how CIM (Convective Interaction Media) monolithic chromatography products solve challenges in DSP for gene therapy and vaccines.

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