Purification of recombinant AAV9 from cell lysates remains a technical challenge and an economic burden for commercial manufacturing of novel gene therapeutics. Currently available affinity resins for AAV9 purification are plagued by poor reduction in Host Cell Protein (HCP) and the inability to use NaOH solutions for Clean-In-Place (CIP). To support the rapidly-expanding gene therapy industry we described the development of AVIPureTM – AAV9, a novel affinity chromatography resin with ability to perform single-step purification of AAV9 from cell lysates with excellent yield and purity. Key distinguishing features are the ability to clean with NaOH and being based on an agarose bead for improved non-specific absorption. The large diameter agarose bead enables high flow rates and excellent resistance to fouling, important features when a typical purification cycle involves loading the column with up to 200 column volumes of host cell culture fluid. The order of magnitude improvement in resin lifetime promises to dramatically decrease resin costs for AAV programs requiring annual production of very large amounts of capsids. The affinity ligand is a novel peptide with exquisite specificity for rAAV9, binding the virus with picomolar affinity. A long, flexible spacer molecule attaches the ligand to the resin, enabling efficient capture of virus. Elution of several variants of AAV9 capsids elution was achieved in the range pH 4.5 – 3. Overall, AVIPureTM – AAV9 resin provides excellent yield, purity and throughput for single-step purification of rAAV9 and enables the use of sodium hydroxide as a cleaning agent for multiple cycles.

Development of AVIPureTM - AAV9: The first affinity resin for AAV production that can be cleaned with sodium hydroxide enables high resin reusability and decreases annual resin costs for high scale production

ACS Fall 2020 Virtual Meeting

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