Mechanistic modeling of empty-full separation in recombinant adeno-associated virus production using anion-exchange membrane chromatography
Gomis-Fons, Joaquin LU ; Zee, Bryan ; Hurwit, Daniel ; Woo, James ; Moscariello, John and Nilsson, Bernt LU
(2024)
In Biotechnology and Bioengineering
121(2).
p.719-734
- Abstract
Recombinant adeno-associated viral vectors (rAAVs) have become an industry-standard technology in the field of gene therapy, but there are still challenges to be addressed in their biomanufacturing. One of the biggest challenges is the removal of capsid species other than that which contains the gene of interest. In this work, we develop a mechanistic model for the removal of empty capsids—those that contain no genetic material—and enrichment of full rAAV using anion-exchange membrane chromatography. The mechanistic model was calibrated using linear gradient experiments, resulting in good agreement with the experimental data. The model was then applied to optimize the purification process through maximization of yield studying the... (More)
Recombinant adeno-associated viral vectors (rAAVs) have become an industry-standard technology in the field of gene therapy, but there are still challenges to be addressed in their biomanufacturing. One of the biggest challenges is the removal of capsid species other than that which contains the gene of interest. In this work, we develop a mechanistic model for the removal of empty capsids—those that contain no genetic material—and enrichment of full rAAV using anion-exchange membrane chromatography. The mechanistic model was calibrated using linear gradient experiments, resulting in good agreement with the experimental data. The model was then applied to optimize the purification process through maximization of yield studying the impact of mobile phase salt concentration and pH, isocratic wash and elution length, flow rate, percent full (purity) requirement, loading density (challenge), and the use of single-step or two-step elution modes. A solution from the optimization with purity of 90% and recovery yield of 84% was selected and successfully validated, as the model could predict the recovery yield with remarkable fidelity and was able to find process conditions that led to significant enrichment. This is, to the best of our knowledge, the first case study of the application of de novo mechanistic modeling for the enrichment of full capsids in rAAV manufacturing, and it serves as demonstration of the potential of mechanistic modeling in rAAV process development.
(Less)
- author
- Gomis-Fons, Joaquin
LU
; Zee, Bryan
; Hurwit, Daniel
; Woo, James
; Moscariello, John
and Nilsson, Bernt
LU
- organization
- publishing date
- 2024-02
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- adeno-associated virus, downstream process development, empty-full separation, full capsid enrichment, mechanistic modeling, membrane chromatography
- in
- Biotechnology and Bioengineering
- volume
- 121
- issue
- 2
- pages
- 16 pages
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:85176265964
- pmid:37942560
- ISSN
- 0006-3592
- DOI
- 10.1002/bit.28595
- language
- English
- LU publication?
- yes
- id
- 61b5e975-d850-4c37-95a6-88019171f2fe
- date added to LUP
- 2024-01-11 09:52:08
- date last changed
- 2025-07-05 23:28:22
@article{61b5e975-d850-4c37-95a6-88019171f2fe,
abstract = {{<p>Recombinant adeno-associated viral vectors (rAAVs) have become an industry-standard technology in the field of gene therapy, but there are still challenges to be addressed in their biomanufacturing. One of the biggest challenges is the removal of capsid species other than that which contains the gene of interest. In this work, we develop a mechanistic model for the removal of empty capsids—those that contain no genetic material—and enrichment of full rAAV using anion-exchange membrane chromatography. The mechanistic model was calibrated using linear gradient experiments, resulting in good agreement with the experimental data. The model was then applied to optimize the purification process through maximization of yield studying the impact of mobile phase salt concentration and pH, isocratic wash and elution length, flow rate, percent full (purity) requirement, loading density (challenge), and the use of single-step or two-step elution modes. A solution from the optimization with purity of 90% and recovery yield of 84% was selected and successfully validated, as the model could predict the recovery yield with remarkable fidelity and was able to find process conditions that led to significant enrichment. This is, to the best of our knowledge, the first case study of the application of de novo mechanistic modeling for the enrichment of full capsids in rAAV manufacturing, and it serves as demonstration of the potential of mechanistic modeling in rAAV process development.</p>}},
author = {{Gomis-Fons, Joaquin and Zee, Bryan and Hurwit, Daniel and Woo, James and Moscariello, John and Nilsson, Bernt}},
issn = {{0006-3592}},
keywords = {{adeno-associated virus; downstream process development; empty-full separation; full capsid enrichment; mechanistic modeling; membrane chromatography}},
language = {{eng}},
number = {{2}},
pages = {{719--734}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Biotechnology and Bioengineering}},
title = {{Mechanistic modeling of empty-full separation in recombinant adeno-associated virus production using anion-exchange membrane chromatography}},
url = {{http://dx.doi.org/10.1002/bit.28595}},
doi = {{10.1002/bit.28595}},
volume = {{121}},
year = {{2024}},
}