Universal Method for the Purification of Recombinant AAV Vectors of Differing Serotypes
| Autor: | Maryellen Mattingly, Denise Woodcock, Jeffery Ardinger, Shelley Nass, Seng H. Cheng, Brenda Burnham, Catherine R. O'Riordan, Shayla E. Osmond, Amy Frederick, Abraham Scaria |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Serotype lcsh:QH426-470 epitope swapping Genetic enhancement viruses Computational biology Article law.invention 03 medical and health sciences 0302 clinical medicine law Genetics Vector (molecular biology) Purification methods lcsh:QH573-671 Molecular Biology Chemistry lcsh:Cytology Transfection Method development lcsh:Genetics 030104 developmental biology Capsid 030220 oncology & carcinogenesis Recombinant DNA Molecular Medicine AAV purification analytical ultracentrifugation |
| Zdroj: | Molecular Therapy: Methods & Clinical Development, Vol 9, Iss, Pp 33-46 (2018) Molecular Therapy. Methods & Clinical Development |
| ISSN: | 2329-0501 |
| Popis: | The generation of clinical good manufacturing practices (GMP)-grade adeno-associated virus (AAV) vectors requires purification strategies that support the generation of vectors of high purity, and that exhibit a good safety and efficacy profile. To date, most reported purification schemas are serotype dependent, requiring method development for each AAV gene therapy product. Here, we describe a platform purification process that is compatible with the purification of multiple AAV serotypes. The method generates vector preparations of high purity that are enriched for capsids with full vector genomes, and that minimizes the fractional content of empty capsids. The two-column purification method, a combination of affinity and ion exchange chromatographies, is compatible with a range of AAV serotypes generated by either the transient triple transfection method or the more scalable producer cell line platform. In summary, the adaptable purification method described can be used for the production of a variety of high-quality AAV vectors suitable for preclinical testing in animal models of diseases. Keywords: AAV purification, epitope swapping, analytical ultracentrifugation |
| Databáze: | OpenAIRE |
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