@article{shastry_barbieri_minzoni_chu_johnson_stoops_pancorbo_gilleskie_ritola_crapanzano_et al._2024, title={Serotype-agnostic affinity purification of adeno-associated virus (AAV) via peptide-functionalized chromatographic resins}, volume={1734}, ISSN={["1873-3778"]}, url={https://doi.org/10.1016/j.chroma.2024.465320}, DOI={10.1016/j.chroma.2024.465320}, journal={JOURNAL OF CHROMATOGRAPHY A}, author={Shastry, Shriarjun and Barbieri, Eduardo and Minzoni, Arianna and Chu, Wenning and Johnson, Stephanie and Stoops, Mark and Pancorbo, Jennifer and Gilleskie, Gary and Ritola, Kimberly and Crapanzano, Michael S. and et al.}, year={2024}, month={Oct} }
 @article{shastry_chu_barbieri_greback-clarke_smith_cummings_minzoni_pancorbo_gilleskie_ritola_et al._2023, title={Rational design and experimental evaluation of peptide ligands for the purification of adeno-associated viruses via affinity chromatography}, volume={9}, ISSN={["1860-7314"]}, DOI={10.1002/biot.202300230}, abstractNote={AbstractAdeno‐associated viruses (AAVs) have acquired a central role in modern medicine as delivery agents for gene therapies targeting rare diseases. While new AAVs with improved tissue targeting, potency, and safety are being introduced, their biomanufacturing technology is lagging. In particular, the AAV purification pipeline hinges on protein ligands for the affinity‐based capture step. While featuring excellent AAV binding capacity and selectivity, these ligands require strong acid (pH <3) elution conditions, which can compromise the product's activity and stability. Additionally, their high cost and limited lifetime has a significant impact on the price tag of AAV‐based therapies. Seeking to introduce a more robust and affordable affinity technology, this study introduces a cohort of peptide ligands that (i) mimic the biorecognition activity of the AAV receptor (AAVR) and anti‐AAV antibody A20, (ii) enable product elution under near‐physiological conditions (pH 6.0), and (iii) grant extended reusability by withstanding multiple regenerations. A20‐mimetic CYIHFSGYTNYNPSLKSC and AAVR‐mimetic CVIDGSQSTDDDKIC demonstrated excellent capture of serotypes belonging to distinct clones/clades – namely, AAV1, AAV2, AAV5, AAV6, AAV8, and AAV9. This corroborates the in silico models documenting their ability to target regions of the viral capsid that are conserved across all serotypes. CVIDGSQSTDDDKIC‐Toyopearl resin features binding capacity (≈1014 vp mL−1) and product yields (≈60%–80%) on par with commercial adsorbents, and purifies AAV2 from HEK293 and Sf9 cell lysates with high recovery (up to 78%), reduction of host cell proteins (up to 700‐fold), and high transduction activity (up to 65%).}, journal={BIOTECHNOLOGY JOURNAL}, author={Shastry, Shriarjun and Chu, Wenning and Barbieri, Eduardo and Greback-Clarke, Paul and Smith, William K. and Cummings, Christopher and Minzoni, Arianna and Pancorbo, Jennifer and Gilleskie, Gary and Ritola, Kimberly and et al.}, year={2023}, month={Sep} }
 @article{kilgore_minzoni_shastry_smith_barbieri_wu_lebarre_chu_o'brien_menegatti_2023, title={The downstream bioprocess toolbox for therapeutic viral vectors}, volume={1709}, ISSN={["1873-3778"]}, DOI={10.1016/j.chroma.2023.464337}, abstractNote={Viral vectors are poised to acquire a prominent position in modern medicine and biotechnology owing to their role as delivery agents for gene therapies, oncolytic agents, vaccine platforms, and a gateway to engineer cell therapies as well as plants and animals for sustainable agriculture. The success of viral vectors will critically depend on the availability of flexible and affordable biomanufacturing strategies that can meet the growing demand by clinics and biotech companies worldwide. In this context, a key role will be played by downstream process technology: while initially adapted from protein purification media, the purification toolbox for viral vectors is currently undergoing a rapid expansion to fit the unique biomolecular characteristics of these products. Innovation efforts are articulated on two fronts, namely (i) the discovery of affinity ligands that target adeno-associated virus, lentivirus, adenovirus, etc.; (ii) the development of adsorbents with innovative morphologies, such as membranes and 3D printed monoliths, that fit the size of viral vectors. Complementing these efforts are the design of novel process layouts that capitalize on novel ligands and adsorbents to ensure high yield and purity of the product while safeguarding its therapeutic efficacy and safety; and a growing panel of analytical methods that monitor the complex array of critical quality attributes of viral vectors and correlate them to the purification strategies. To help explore this complex and evolving environment, this study presents a comprehensive overview of the downstream bioprocess toolbox for viral vectors established in the last decade, and discusses present efforts and future directions contributing to the success of this promising class of biological medicines.}, journal={JOURNAL OF CHROMATOGRAPHY A}, author={Kilgore, Ryan and Minzoni, Arianna and Shastry, Shriarjun and Smith, Will and Barbieri, Eduardo and Wu, Yuxuan and Lebarre, Jacob P. and Chu, Wenning and O'Brien, Juliana and Menegatti, Stefano}, year={2023}, month={Oct} }