Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Optimizing T cell responses of targeted peptide antigen delivery by modulating antigen processing through amino acid exchange.

International journal of biological macromolecules·2026
Same author

Efficiency and scalability in harvesting mammalian cell cultures: A scale-down approach to continuous centrifugation.

Journal of biotechnology·2026
Same author

Antagonist nanobodies prevent protease inhibition by CD109.

The Journal of biological chemistry·2026
Same author

Cell specific nutrient feeding in perfusion mode enhances hMSC growth in stirred tank bioreactor process.

Journal of biotechnology·2025
Same author

Single-cell transcriptomics reveals dynamics of natural killer cell expansion in a feeder cell-free culture of peripheral blood mononuclear cells-implications for immunotherapy.

Cytotherapy·2025
Same author

Heterogeneity in an adeno-associated virus transfection-based production process limits the production efficiency.

Scientific reports·2025

Related Experiment Video

Updated: Mar 7, 2026

Production, Purification, and Quality Control for Adeno-associated Virus-based Vectors
09:21

Production, Purification, and Quality Control for Adeno-associated Virus-based Vectors

Published on: January 29, 2019

38.1K

Automated plasmid purification system enables increased AAV production.

Hannes Thorell1, Sanne Rönning1, Olivér Daoda2

  • 1KTH Royal Institute of Technology, Department of Protein Science, Stockholm SE-106 91, Sweden.

New Biotechnology
|March 5, 2026
PubMed
Summary
This summary is machine-generated.

Automated plasmid DNA purification significantly reduces manufacturing time for adeno-associated virus (AAV) vectors. While yields may decrease, the automated method surprisingly boosts AAV production efficiency in specific systems.

Keywords:
Adeno-associated virusautomationdrug developmentgene therapyplasmidproductionpurification

More Related Videos

Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors
09:12

Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors

Published on: April 5, 2024

4.7K
Process Development for the Production and Purification of Adeno-Associated Virus AAV2 Vector using Baculovirus-Insect Cell Culture System
10:31

Process Development for the Production and Purification of Adeno-Associated Virus AAV2 Vector using Baculovirus-Insect Cell Culture System

Published on: January 13, 2022

10.5K

Related Experiment Videos

Last Updated: Mar 7, 2026

Production, Purification, and Quality Control for Adeno-associated Virus-based Vectors
09:21

Production, Purification, and Quality Control for Adeno-associated Virus-based Vectors

Published on: January 29, 2019

38.1K
Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors
09:12

Author Spotlight: Improved Method for Production and Purification of Adeno-Associated Viral Vectors

Published on: April 5, 2024

4.7K
Process Development for the Production and Purification of Adeno-Associated Virus AAV2 Vector using Baculovirus-Insect Cell Culture System
10:31

Process Development for the Production and Purification of Adeno-Associated Virus AAV2 Vector using Baculovirus-Insect Cell Culture System

Published on: January 13, 2022

10.5K

Area of Science:

  • Biotechnology
  • Gene Therapy Manufacturing
  • Molecular Biology

Background:

  • Adeno-associated virus (AAV) vectors are crucial for gene therapies, but high manufacturing costs hinder development and access.
  • Plasmid DNA (pDNA) purification is a significant bottleneck in AAV production, especially at larger scales.
  • Current manual plasmid purification methods are time-consuming and limit scalability.

Purpose of the Study:

  • To compare the efficiency and outcomes of manual versus automated plasmid DNA purification systems for AAV production.
  • To evaluate the impact of scale (Maxi, Mega, Giga) on purification time, yield, and endotoxin levels.
  • To investigate the underlying mechanisms for differences in AAV production titres observed between purification methods.

Main Methods:

  • Plasmid DNA purification using conventional manual kits and a scalable automated system at Maxi, Mega, and Giga scales.
  • AAV vector production via PEI-based transfection of HEK293F cells using purified pDNA.
  • Analysis of AAV9 titres, endotoxin levels, and pDNA complex formation using dynamic light scattering.

Main Results:

  • Automated purification significantly reduced processing time, with savings increasing at larger scales (e.g., 8 hours saved at Giga scale).
  • The automated system generally resulted in lower yields (3-67% reduction) and higher endotoxin levels compared to manual methods.
  • Surprisingly, plasmids purified by the automated system led to a 14-fold increase in AAV-MAX production titres, attributed to larger pDNA transfection complexes formed by the automated system's elution buffer.

Conclusions:

  • Automated plasmid purification offers substantial time savings for AAV manufacturing, particularly at scale.
  • The choice of purification method and its associated elution buffer can critically impact transfection efficiency and subsequent biologics production.
  • Optimization of plasmid preparation methods, including elution buffer composition, holds potential for improving transient production of AAV and other biologics.