Model-based process optimization for mAb chromatography

Kozorog, Mirijam; Caserman, Simon; Grom, Matic; Vicente, Filipa A.; Pohar, Andrej; Likozar, Blaž

doi:10.1016/j.seppur.2022.122528

Cited by 7 publications

(1 citation statement)

References 30 publications

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“…The recent consolidation of an industrial layout for manufacturing of adeno‐associated viruses (AAVs)—inspired by the downstream process platform established for monoclonal antibodies (Gao et al, 2020; Kozorog et al, 2023; Matos et al, 2021; Zhang et al, 2015)—highlights the role of affinity chromatography as the keystone in the purification of viral vectors for cell and gene therapies (Fortuna et al, 2019; Moleirinho et al, 2020). For AAVs, a number of affinity adsorbents are available: POROS™ CaptureSelect™ AAVX and AVB Sepharose HP are commercially available as pan‐selective resins, while CaptureSelect™ AAV8 and AAV9, and AVIPure® AAV2, AAV8, and AAV9 are available for serotype‐specific applications (Adams et al, 2020; van Lieshout et al, 2023).…”

Section: Introductionmentioning

confidence: 99%

Peptide ligands targeting the vesicular stomatitis virus G (VSV‐G) protein for the affinity purification of lentivirus particles

Barbieri,

Mollica,

Moore

et al. 2023

Biotech & Bioengineering

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The recent uptick in the approval of ex vivo cell therapies highlights the relevance of lentivirus (LV) as an enabling viral vector of modern medicine. As labile biologics, however, LVs pose critical challenges to industrial biomanufacturing. In particular, LV purification—currently reliant on filtration and anion‐exchange or size‐exclusion chromatography—suffers from long process times and low yield of transducing particles, which translate into high waiting time and cost to patients. Seeking to improve LV downstream processing, this study introduces peptides targeting the enveloped protein Vesicular stomatitis virus G (VSV‐G) to serve as affinity ligands for the chromatographic purification of LV particles. An ensemble of candidate ligands was initially discovered by implementing a dual‐fluorescence screening technology and a targeted in silico approach designed to identify sequences with high selectivity and tunable affinity. The selected peptides were conjugated on Poros resin and their LV binding‐and‐release performance was optimized by adjusting the flow rate, composition, and pH of the chromatographic buffers. Ligands GKEAAFAA and SRAFVGDADRD were selected for their high product yield (50%–60% of viral genomes; 40%–50% of HT1080 cell‐transducing particles) upon elution in PIPES buffer with 0.65 M NaCl at pH 7.4. The peptide‐based adsorbents also presented remarkable values of binding capacity (up to 3·109 TU per mL of resin, or 5·1011 vp per mL of resin, at the residence time of 1 min) and clearance of host cell proteins (up to a 220‐fold reduction of HEK293 HCPs). Additionally, GKEAAFAA demonstrated high resistance to caustic cleaning‐in‐place (0.5 M NaOH, 30 min) with no observable loss in product yield and quality.

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