Overcoming the Hurdles of Density Gradient Ultracentrifugation for Optimized Gene Therapy Purification Workflows

Speaker:
Ross VerHeul
Senior Applications Scientist - Beckman Coulter

Abstract:
Centrifugation has long served as a critical separation tool in countless research and production facilities spanning a wide range of disciplines. Modern centrifuges reach incredibly high speeds (with centrifugal acceleration sometimes exceeding 1,000,000 x g) to aid the high-resolution separation of particles, typically in the micro- or nanometer range, by size and/or density. Today’s gene therapy products, such as viral vectors, require high-quality purification to ensure the consistent production of safe, efficacious therapeutics of the highest quality to further advance this rapidly growing field and deliver solutions to patients in need.

Density gradient ultracentrifugation (DGUC) is a centrifuge-based technique for providing superior purification of viral vectors (e.g., isolating full AAV particles from partial and empty capsids), along with other materials (such as plasmid DNA) in gene therapy production workflows. Though a well-established and mature method, DGUC is sometimes viewed as dated, challenging to design and conduct, or only suited for small-scale research applications. In this webinar, we’ll address these perceptions and discuss the premise of DGUC as a high-resolution purification technique for AAVs and plasmid DNA. We’ll also provide guidance on how to get started with DGUC and optimize this technique for gene therapy workflows.

Learning Objectives:

◦ Learn about the various types of density gradients techniques,
  how they are used, and how to get started.

◦ Learn how to design and optimize a density gradient
  ultracentrifugation workflow, with AAV and plasmid DNA
  purification as examples.

◦ Learn how an optimized DGUC procedure can significantly
  increase throughput (by up to 7.5-fold) and become suitable
  for larger-scale workflows