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Single-Use Capture Purification of Adeno-Associated Viral Gene Transfer Vectors by Membrane-Based Steric Exclusion Chromatography

  • Pavel Marichal-Gallardo 1
  • Kathleen Börner 2
  • Michael M. Pieler 1
  • Vera Sonntag-Buck 2
  • Martin Obr 2
  • David Bejarano 2
  • Michael W. Wolff 3
  • Hans-Georg Kräusslich 2
  • Udo Reichl 4
  • Dirk Grimm 2
  • 1 - Bioprocess Engineering - Max Planck Institute for Dynamics of Complex Technical Systems - Magdeburg Germany
  • 2 - Center for Infectious Diseases - Virology - Heidelberg University Hospital - Heidelberg Germany
  • 3 - Institute of Bioprocess Engineering and Pharmaceutical Technology - Technische Hochschule Mittelhessen - Gießen Germany
  • 4 - Bioprocess Engineering - Otto von Guericke University Magdeburg - Magdeburg Germany

Abstract

We present membrane-based steric exclusion chromatography (SXC) as a universal capture step for purification of adeno-associated virus (AAV) gene transfer vectors independent of their serotype and surface characteristics. SXC is performed by mixing an unpurified cell culture supernatant containing AAV particles with polyethylene glycol (PEG) and feeding the mixture onto a chromatography filter unit. The purified AAV particles are recovered by flushing the unit with a solution lacking PEG. SXC is an inexpensive single-use method that permits to concentrate, purify, and re-buffer AAV particles with yields >95% and >80% impurity clearance. SXC could theoretically be employed at industrial scales with units of nearly 20 m2.


 


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