Optimization on cell lysis and capture process of human adenovirus type 5 produced in suspension HEK293 cells

Abstract

Adenovirus (Adv) is increasingly recognized for its significance in the fields of gene therapy and viral vector vaccines. The diverse applications in clinical trials and fundamental research necessitate the development of environmentally and economically sustainable purification processes that are straightforward and scalable for both academic and industrial contexts. In the initial segment of this study, we evaluated the lysis efficiency of polysorbate 20 (PS20) in comparison to polysorbate 80 (PS80). Our findings indicated that the viability HEK293 could be reduced to approximately 13 %, with a detectable Adv5 concentration of average 1.62 × 10⁹ ifu/mL in the supernatant after an incubation in 1.0 % (w/w) PS20 or PS80 buffer. In the subsequent portion of this research, we employed a high-throughput static binding capacity (SBC) screening tool in conjunction with on-column dynamic binding capacity (DBC) validation to concurrently assess the binding capacity of Adv5 on nine different types of anion exchange media. The results demonstrated that both Sartobind Q membrane and POROS 50HQ resin exhibited binding capacities exceeding 1.0 × 10¹³ vp/mL under the testing conditions.