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Related Experiment Video

Updated: Jun 24, 2026

Process Development for the Production and Purification of Adeno-Associated Virus (AAV)2 Vector using Baculovirus-Insect Cell Culture System
10:31

Process Development for the Production and Purification of Adeno-Associated Virus (AAV)2 Vector using Baculovirus-Insect Cell Culture System

Published on: January 13, 2022

Scaleable chromatographic purification process for recombinant adeno-associated virus (rAAV).

C R O'Riordan1, A L Lachapelle, K A Vincent

  • 1Genzyme Corporation, Framingham, MA 01701-9322, USA. coriordan@genzyme.com

The Journal of Gene Medicine
|February 24, 2001
PubMed
Summary
This summary is machine-generated.

A new column chromatography method rapidly purifies recombinant adeno-associated virus (rAAV) vectors. This scalable process yields highly pure, biologically active rAAV, suitable for clinical gene therapy applications.

More Related Videos

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Isolation of Adeno-Associated Viral Vectors Through a Single-Step and Semi-Automated Heparin Affinity Chromatography Protocol
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Last Updated: Jun 24, 2026

Process Development for the Production and Purification of Adeno-Associated Virus (AAV)2 Vector using Baculovirus-Insect Cell Culture System
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Process Development for the Production and Purification of Adeno-Associated Virus (AAV)2 Vector using Baculovirus-Insect Cell Culture System

Published on: January 13, 2022

Suspension Culture Production and Purification of Adeno-Associated Virus by Iodixanol Density Gradient Centrifugation for In Vivo Applications
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Suspension Culture Production and Purification of Adeno-Associated Virus by Iodixanol Density Gradient Centrifugation for In Vivo Applications

Published on: February 9, 2024

Isolation of Adeno-Associated Viral Vectors Through a Single-Step and Semi-Automated Heparin Affinity Chromatography Protocol
09:12

Isolation of Adeno-Associated Viral Vectors Through a Single-Step and Semi-Automated Heparin Affinity Chromatography Protocol

Published on: April 5, 2024

Area of Science:

  • Molecular Biology
  • Virology
  • Biotechnology

Background:

  • Adeno-associated virus (AAV) is a human parvovirus utilized in gene therapy.
  • Traditional purification methods like CsCl gradients are unsuitable for large-scale manufacturing and yield contaminated products.
  • A need exists for efficient, scalable purification of recombinant AAV (rAAV) vectors.

Purpose of the Study:

  • To develop a process-scale method for rapid and efficient purification of rAAV from cellular lysates.
  • To establish a scalable purification strategy for clinical-grade rAAV vectors.

Main Methods:

  • Purification involves treating infected cell lysates with trypsin and nuclease.
  • Utilizes a combination of ion exchange chromatography (ceramic hydroxyapatite, DEAE-Sepharose) and affinity chromatography (cellufine sulphate).
  • Exclusively employs column chromatography for purification from crude cellular lysates co-infected with adenovirus.

Main Results:

  • Achieved purification of rAAV particles solely through column chromatography.
  • Demonstrated >90% purity of column-purified rAAV.
  • Confirmed rAAV was free of detectable adenovirus contamination, biologically active, and capable of efficient gene transfer in vivo.

Conclusions:

  • Column chromatography alone is feasible for isolating highly purified rAAV vectors.
  • The described methods represent advancements in rAAV purification.
  • These purification procedures are adaptable for commercial production of clinical-grade rAAV vectors.