학술논문
Single-Use Capture Purification of Adeno-Associated Viral Gene Transfer Vectors by Membrane-Based Steric Exclusion Chromatography.
Document Type
Article
Author
Source
Subject
*GENETIC vectors
*GENETIC transformation
*VIRAL genes
*CHROMATOGRAPHIC analysis
*POLYETHYLENE glycol
*ADENO-associated virus
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Language
ISSN
1043-0342
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. [ABSTRACT FROM AUTHOR]