Design space determination to optimize DNA complexation and full capsid formation in transient rAAV manufacturing
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Le résumé fourni par la source
Recombinant adeno-associated virus (rAAV) vectors are a promising platform for in vivo gene therapies. However, cost-effective, well-characterized processes necessary to manufacture rAAV therapeutics are challenging to develop without an understanding of how process parameters (PPs) affect rAAV product quality attributes (PQAs). In this work, a central composite orthogonal experimental design was employed to examine the influence of four PPs for transient transfection complex formation (polyethylenimine:DNA [PEI:DNA] ratio, total DNA/cell, cocktail volume, and incubation time) on three rAAV PQAs related to capsid content (vector genome titer, vector genome:capsid particle ratio, and two-dimensional vector genome titer ratio). A regression model was established for each PQA using partial least squares, and a design space (DS) was defined in which Monte Carlo simulations predicted < 1% probability of failure (POF) to meet predetermined PQA specifications. Of the three PQAs, viral genome titer was most strongly correlated with changes in complexation PPs. The DS and acceptable PP ranges were largest when incubation time and cocktail volume were kept at mid-high setpoints, and PEI:DNA ratio and total DNA/cell were at low-mid setpoints. Verification experiments confirmed model predictive capability, and this work establishes a framework for studying other rAAV PPs and their relationship to PQAs.
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Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- Design space determination to optimize DNA complexation and full capsid formation in transient rAAV manufacturing
- Date Crossref
- 21/07/2023
- Éditeur
- Wiley
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
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University of Massachusetts Lowell Department of Biomedical Engineering and Biotechnology pays non établi dans la noticeUniversité ou école supérieure
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University of Delaware Department of Chemical and Biomolecular Engineering pays non établi dans la noticeUniversité ou école supérieure
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Johns Hopkins University Department of Chemical and Biomolecular Engineering pays non établi dans la noticeUniversité ou école supérieure
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University of Massachusetts Chan Medical School MassBiologics pays non établi dans la noticeUniversité ou école supérieure
Department of Biomedical Engineering and Biotechnology — University of Massachusetts Lowell, Department of Chemical and Biomolecular Engineering — University of Delaware et Department of Chemical and Biomolecular Engineering — Johns Hopkins University, avec 1 autre affiliation.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.