Achieve significantly increased adenovirus yield with density gradient ultracentrifugation: a comparative study

Live30 webinars are thirty minute presentations designed to update you on the latest innovations, applications and data in a fast yet interactive format.

Adenoviral (AdV) vectors are highly effective tools for gene therapy (GT), oncolytic virotherapy, and vaccines due to their high gene transduction efficiencies, safety, and tunability. Although numerous technologies exist for the downstream purification of AdV, several critical manufacturing challenges still exist -- including scalability, low recovery, and the removal of highly similar impurities such as empty capsids.

Two of the most prominent AdV purification techniques include density gradient ultracentrifugation (DGUC) and ion-exchange chromatography (IEC). Although IEC is well established as a scalable approach to biomolecule purification with short separation times, it suffers from several limitations in the context of AdV, including low binding capacity, serotype dependency, and a tradeoff between yield and purity. DGUC, however, is a robust, serotype-independent method that offers improved product recovery by exploiting subtle differences in the buoyant density between full, therapeutically active AdV particles and other process impurities.

Here, we directly compare both techniques and demonstrate significant improvements in product critical quality attributes (CQAs) including >200-fold increases in yield and concentration for the DGUC-purified AdV5. In addition, we will discuss approaches to maximize throughput and efficiency when scaling up or down, which are universally applicable across a range of therapeutic platforms such as AdV, adeno-associated virus (AAV), and more.

Watch this webinar to learn about:

• How DGUC and IEC perform with respect to various AdV CQAs
• Key considerations for choosing DGUC or IEC for AdV purification
• Solutions for rapid implementation, optimization, and scaling of DGUC for purification

Shawn Sternisha PhD

Senior Field Applications Scientist, Beckman Coulter

Shawn Sternisha is currently a Senior Field Applications Scientist in the Biotechnology Business Unit at Beckman Coulter Life Sciences where he develops applications and supports centrifuge product lines. Shawn attended Illinois State University for his undergraduate studies and went on to earn his Ph.D. in Biochemistry at Florida State University. His doctoral research was focused on understanding how glucokinase regulates glucose homeostasis at the molecular level. Shawn has also previously worked at GlaxoSmithKline where he was involved in investigating novel imaging modalities for monitoring mammalian cell cultures. Shawn is broadly interested in enzyme structure-function relationships and mechanisms of action, cell and gene therapy, and expanding applications of biochemical and biophysical techniques (namely Analytical Ultracentrifugation). Shawn currently resides in Miami, FL with his fiancée Patty and their dog Ruby. In his free time, he enjoys cooking, running, traveling, and learning Spanish.

Watch now

SPEAKERS

Shawn Sternisha PhD

Senior Field Applications Scientist, Beckman Coulter

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