Multimodal production of adeno-associated virus
Document Type
Article
Abstract
Adeno-associated virus (AAV) is an increasingly popular tool in the research laboratory, and use of this viral vector clinically is occurring at an accelerated pace. Nevertheless, despite its popularity, AAV is a relatively cumbersome virus to produce; however, significant efforts have been invested to develop, optimize, and simplify methodology that allows the generation of high-quality AAV with significantly increased production yields. Here we describe multiple modalities for production and purification of AAV particles produced in HEK293 cell cultures using an iodixanol density gradient. We include two methods adapted for harvesting virus from the culture media: Tangential flow filtration (TFF) and polyethylene glycol precipitation (PEGylation). Moreover, we also describe the protocol for anion exchange chromatography, which can be used after the iodixanol gradient as an additional purification step. Last, we provide various protocols for determining virus titer.
Keywords
AAV, AAV production, Digital droplet PCR, Iodixanol gradient, PEGylation, Tangential flow filtration
Publication Date
1-1-2019
Publication Title
Methods in Molecular Biology
ISSN
10643745
Volume
1937
First Page
101
Last Page
124
PubMed ID
30706392
Digital Object Identifier (DOI)
10.1007/978-1-4939-9065-8_6
Recommended Citation
Sandoval, Ivette M.; Kuhn, Nathan M.; and Manfredsson, Fredric P., "Multimodal production of adeno-associated virus" (2019). Translational Neuroscience. 2031.
https://scholar.barrowneuro.org/neurobiology/2031