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Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin
Recent successes of adeno-associated virus (AAV)–based gene therapy have created a demand for large-scale AAV vector manufacturing and purification techniques for use in clinical trials and beyond. During the development of purification protocols for rh.10, hu.37, AAV8, rh.64R1, AAV3B, and AAV9 vect...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4632836/ https://www.ncbi.nlm.nih.gov/pubmed/26605372 http://dx.doi.org/10.1038/mtm.2015.40 |
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author | Wang, Qiang Lock, Martin Prongay, Andrew J Alvira, Mauricio R Petkov, Boris Wilson, James M |
author_facet | Wang, Qiang Lock, Martin Prongay, Andrew J Alvira, Mauricio R Petkov, Boris Wilson, James M |
author_sort | Wang, Qiang |
collection | PubMed |
description | Recent successes of adeno-associated virus (AAV)–based gene therapy have created a demand for large-scale AAV vector manufacturing and purification techniques for use in clinical trials and beyond. During the development of purification protocols for rh.10, hu.37, AAV8, rh.64R1, AAV3B, and AAV9 vectors, based on a widely used affinity resin, AVB sepharose (GE), we found that, under the same conditions, different serotypes have different affinities to the resin, with AAV3B binding the best and AAV9 the poorest. Further analysis revealed a surface-exposed residue (amino acid number 665 in AAV8 VP1 numbering) differs between the high-affinity AAV serotypes (serine in AAV3B, rh.10, and hu.37) and the low-affinity ones (asparagine in AAV8, rh.64R1, and AAV9). The residue locates within a surface-exposed, variable epitope flanked by highly conserved residues. The substitution of the epitope in AAV8, rh.64R1, and AAV9 with the corresponding epitope of AAV3B (SPAKFA) resulted in greatly increased affinity to AVB sepharose with no reduction in the vectors’ in vitro potency. The presence of the newly identified AVB-binding epitope will be useful for affinity resin selection for the purification of novel AAV serotypes. It also suggests the possibility of vector engineering to yield a universal affinity chromatography purification method for multiple AAV serotypes. |
format | Online Article Text |
id | pubmed-4632836 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-46328362015-11-24 Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin Wang, Qiang Lock, Martin Prongay, Andrew J Alvira, Mauricio R Petkov, Boris Wilson, James M Mol Ther Methods Clin Dev Article Recent successes of adeno-associated virus (AAV)–based gene therapy have created a demand for large-scale AAV vector manufacturing and purification techniques for use in clinical trials and beyond. During the development of purification protocols for rh.10, hu.37, AAV8, rh.64R1, AAV3B, and AAV9 vectors, based on a widely used affinity resin, AVB sepharose (GE), we found that, under the same conditions, different serotypes have different affinities to the resin, with AAV3B binding the best and AAV9 the poorest. Further analysis revealed a surface-exposed residue (amino acid number 665 in AAV8 VP1 numbering) differs between the high-affinity AAV serotypes (serine in AAV3B, rh.10, and hu.37) and the low-affinity ones (asparagine in AAV8, rh.64R1, and AAV9). The residue locates within a surface-exposed, variable epitope flanked by highly conserved residues. The substitution of the epitope in AAV8, rh.64R1, and AAV9 with the corresponding epitope of AAV3B (SPAKFA) resulted in greatly increased affinity to AVB sepharose with no reduction in the vectors’ in vitro potency. The presence of the newly identified AVB-binding epitope will be useful for affinity resin selection for the purification of novel AAV serotypes. It also suggests the possibility of vector engineering to yield a universal affinity chromatography purification method for multiple AAV serotypes. Nature Publishing Group 2015-11-04 /pmc/articles/PMC4632836/ /pubmed/26605372 http://dx.doi.org/10.1038/mtm.2015.40 Text en Copyright © 2015 Official journal of the American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/ |
spellingShingle | Article Wang, Qiang Lock, Martin Prongay, Andrew J Alvira, Mauricio R Petkov, Boris Wilson, James M Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin |
title | Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin |
title_full | Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin |
title_fullStr | Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin |
title_full_unstemmed | Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin |
title_short | Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin |
title_sort | identification of an adeno-associated virus binding epitope for avb sepharose affinity resin |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4632836/ https://www.ncbi.nlm.nih.gov/pubmed/26605372 http://dx.doi.org/10.1038/mtm.2015.40 |
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