Downstream Process Development for an Adeno-associated Virus Generated from HeLa Cell Line
View/ Open
Hanson Zhou Thesis.pdf (1.201Mb)
Access Status
Full text of the requested work is not available in DASH at this time ("restricted access"). For more information on restricted deposits, see our FAQ.Author
Zhou, Hanson
Metadata
Show full item recordCitation
Zhou, Hanson. 2022. Downstream Process Development for an Adeno-associated Virus Generated from HeLa Cell Line. Master's thesis, Harvard University Division of Continuing Education.Abstract
The goal of this work was to assemble a downstream purification process foradeno-associated virus (AAV) gene therapy product utilizing a HeLa cell line in
Ultragenyx. HeLa cells were grown by upstream team and transfected with AAV.
Harvest of cells resulted in gaining the intended gene therapy product, as well as several
impurities that must be reduced or eliminated for drug safety. Additionally, an efficacious
product must be enriched and concentrated. The downstream process consisted of several
distinct unit operations to achieve these goals. Several platform processes and materials
were adapted and evaluated, in addition to several new techniques, to develop a scalable
downstream process. Clarification resulted in a new filter to be used, while AEX
membrane filtration utilized a preexisting filter. Affinity chromatography also resulted in
platform chromatography being used. Post heat inactivation, and after AEX
chromatography resin selection, downstream unit operations resulted in a successful
purification of this gene therapy product. Additionally, several unit operations yielded
interesting findings that are then discussed, such as load conductivity impacts during
clarification. The overall downstream process is discussed holistically, and suggestions
are made for further possible improvements of this gene therapy product, with the hope
of improving other AAV products within Ultragenyx.
Terms of Use
This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAACitable link to this page
https://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37372497
Collections
- DCE Theses and Dissertations [1331]
Contact administrator regarding this item (to report mistakes or request changes)