Adenovirus purification by two-column, size-exclusion, simulated countercurrent chromatography

Nestola, Piergiuseppe; Silva, Ricardo J.S.; Peixoto, Cristina; Alves, Paula M.; Carrondo, Manuel J.T.; Mota, José P. B.

doi:10.1016/j.chroma.2014.04.079

Cited by 49 publications

(35 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To date, biopharmaceuticals targeted by SMB chromatography (SMBC) are mainly proteins and mAbs. Nevertheless, SEC-SMBC has been recently applied to purification of cell culture-derived IAV [34] and human AdV type 5 (AdV-5) [35] using columns packed with Seph 4 FF resin. Particularly promising results were obtained by Nestola et al [35] with a two-column setup yielding higher virus recovery and purity (HC DNA and protein clearance) than the batch (single-column) SEC process with the same amount of stationary phase.…”

Section: Size Exclusion Chromatography (Sec)mentioning

confidence: 99%

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

Junter

Lebrun

2020

Journal of Pharmaceutical Analysis

View full text Add to dashboard Cite

Section: Size Exclusion Chromatography (Sec)mentioning

confidence: 99%

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

Junter

Lebrun

2020

Journal of Pharmaceutical Analysis

View full text Add to dashboard Cite

“…Previous reports showed that the CIM™ QA-1 was preferable over the weak anion CIM™ DEAE-1. Nestola, Silva, et al (2014) described the purification of Ad5 using a two column, quasicontinuous, simulated moving-bed size exclusion chromatography (SEC) process which achieved a recovery of 86% determined by real-time polymerase chain reaction (PCR). Another work using a porous cast membrane Peixoto, Ferreira, Sousa, Carrondo, and Alves (2008) achieved a 62% recovery (determined by cell fluorescence) of infectious Ad5.…”

Section: Introductionmentioning

confidence: 99%

Adenovirus 5 recovery using nanofiber ion‐exchange adsorbents

Turnbull

Wright

Green

et al. 2019

Biotech & Bioengineering

View full text Add to dashboard Cite

Viral vectors such as adenovirus have successful applications in vaccines and gene therapy but the manufacture of the high-quality virus remains a challenge. It is desirable to use the adsorption-based chromatographic separations that so effectively underpin the therapeutic protein manufacture. However fundamental differences in the size and stability of this class of product mean it is necessary to revisit the design of sorbent's morphology and surface chemistry. In this study, the behaviour of a cellulose nanofiber ion-exchange sorbent derivatised with quaternary amine ligands at defined densities is characterised to address this. This material was selected as it has a large accessible surface area for viral particles and rapid process times. Initially, the impact of surface chemistry on infective product recovery using low (440 µmol/g), medium (750 µmol/g), and high (1029 µmol/g) ligand densities is studied. At higher densities product stability is reduced, this effect increased with prolonged adsorption durations of 24 min with just~10% loss at low ligand density versus~50% at high. This could be mitigated by using a high flow rate to reduce the cycle time to~1 min. Next, the impact of ligand density on the separation's resolution was evaluated. Key to understanding virus quality is the virus particle: infectious virus particle ratio. It was found this parameter could be manipulated using ligand density and elution strategy. Together this provides a basis for viral vector separations that allows for their typically low titres and labile nature by using high liquid velocity to minimise both load and on-column times while separating key product and process-related impurities.

show abstract

“…Warikoo et al provided the proof of concept for the successful integration of a perfusion bioreactor with a continuous capture step, and this group demonstrated the first fully continuous end‐to‐end production of a monoclonal antibody that integrated a perfusion bioreactor and the downstream operations of affinity capture, viral inactivation, and two ion exchange polishing steps . In recent years, several other groups reported successful strategies for continuous bioprocessing . In addition, modeling studies underlined the potential for economic benefit and delineated decisive factors for significant production cost savings associated with continuous downstream processing …”

Section: Introductionmentioning

confidence: 99%

Evaluation of continuous nonaffinity capture chromatography for a recombinant enzyme—optimization for a changing perfusion feedstream and comparison to batch processing

Thompson¹,

Wilson²,

Larkin

et al. 2018

Biotechnology Progress

View full text Add to dashboard Cite

This work presents the optimization and critical evaluation of continuous capture chromatography in the downstream process of a recombinant enzyme. For the upstream manufacturing of this molecule, a perfusion process was implemented due to benefits for product quality and productivity. This process is, however, characterized by low titer and significant changes over the course of the harvest duration in terms of active enzyme concentration and impurity content. We evaluated the feasibility and benefits of a continuous capture operation. This case study illustrates the design approach that can be utilized to address challenges presented by a changing feedstream, and the statistical measures that can be employed to characterize and optimize the operating space under material and time constraints. Process economic modeling in conjunction with Monte Carlo simulations indicate that even for a nonaffinity capture step utilizing a relatively cheap ion-exchange resin, the smaller column volume used in a continuous set-up results in cost savings compared to the batch process. We compare this option to the scenario of repeated processing using a small capture column in batch mode. Our analysis establishes that continuous processing becomes economically attractive for processes where only a small portion of the potential column lifetime can be utilized or for column steps with slow mass transport and shallow breakthrough curves. In cases where column breakthrough is sharp and resin lifetime is relatively short, continuous processing may offer an improvement over traditional batch processing, but much of the productivity and cost savings can be realized through repeated column cycling. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018 © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1195-1204, 2018.

show abstract

Adenovirus purification by two-column, size-exclusion, simulated countercurrent chromatography

Cited by 49 publications

References 38 publications

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

Adenovirus 5 recovery using nanofiber ion‐exchange adsorbents

Evaluation of continuous nonaffinity capture chromatography for a recombinant enzyme—optimization for a changing perfusion feedstream and comparison to batch processing

Contact Info

Product

Resources

About