11/10/2022
By Seongkyu Yoon

Zuckerberg College of Health Sciences, Department of Biomedical and Nutritional Sciences, invites you to attend a doctoral dissertation proposal defense in Pharmaceutical Sciences by Yong Suk Lee on “A complete quantification of full, partial, and empty capsid in rAAV products via a fluorescence-based Multicolumn countercurrent solvent gradient purification (MCSGP) IEX-HPLC Method.”

Date: Monday, Nov. 21, 2022
Time: 1 to 3 p.m.
Location: University Crossing 415

Committee:

  • Jonghan Kim, Associate Professor, Biomedical & Nutritional Sciences, University of Massachusetts Lowell
  • Shannon L. Kelleher, Professor, Biomedical & Nutritional Sciences, University of Massachusetts Lowell
  • David McNally, Assistant Professor, Medicine, UMass Chan Medical School
  • Raghu Shivappa, Vice President, Global Biologics and Gene Therapy Process Development, Takeda

Abstract:
The recombinant adeno-associated virus (rAAV) has shown its great potential as a viral vector in gene therapy due to its low immunogenicity, minimized pathogenicity, and capability to target different tissues depending on different serotypes. It is a gold standard to produce rAAVs via triple transfection. Due to imperfect packaging efficiency and a lack of understanding of encapsidation processes, inevitably, there is a population of empty, partial, and other capsids formed during manufacturing. This product-related impurity provokes an immune response without any therapeutic effect when injected into a patient. Therefore, a simple, rapid, and sample-sparing method to determine the full and empty ratio of the samples across the manufacturing process is needed to monitor and control the quality of gene therapy-related products.
The proposed effort aims to elucidate accurate measurement of capsid ratio by utilizing multicolumn countercurrent solvent gradient purification (MCSGP) in both anion and cation environments under fluorescence detection to reach baseline separation of empty and partial from full capsids. Moreover, the appropriate concentration of divalent cations and surfactants will be explored to improve the on-column stability of AAV for possible heat stress and aggregation, respectively.