Cell and gene therapies (CGTs) are at the forefront of innovation to treat severe diseases, such as cancer, as well as rare diseases.<b> </b>To support the manufacturing of Cell and Gene Therapies, we have developed innovative chromatography technologies, specifically designed to improve the downstream purification of viral vectors, such as the target Adeno-Associated Virus (AAV). This work represents the development of short synthetic AAV-binding peptides and demonstrates their applicability as affinity ligands for the purification of AAV from HEK cell culture harvests. Candidate AAV-targeting peptides were initially identified by screening a solid-phase peptide library against a mixture of AAV spiked in HEK host cell proteins through microfluidic screening technology. An ensemble of homologous sequences was identified, conjugated on Toyopearl resin, and evaluated by the purification performance of AAV from HEK cell culture harvests. Notably, four potential sequences like FWNWHHFK, show the potential to isolate target AAV from HEK cell culture harvests with high affinity and selectivity, which could be a significant improvement to the downstream process of viral vectors, by reducing the purification steps and maximizing productivity.

High-throughput screening of peptide ligands for the purification of Adeno-associated virus from HEK cell culture harvests

ACS Spring 2022

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