Recovery of infective virus particles in ion-exchange and hydrophobic interaction monolith chromatography is influenced by particle charge and total-to-infective particle ratio

Sviben, Dora; Forčić, Dubravko; Ivancic-Jelecki, Jelena; Halassy, Beata; Brgles, Marija

doi:10.1016/j.jchromb.2017.04.015

Cited by 30 publications

(32 citation statements)

References 36 publications

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“…Regulatory agencies established a limit of 10 ng of dsDNA and 100 ppm of HCP per dose in the final biological product (Eaton, 1995; Knezevic, Stacey, & Petricciani, 2008). However, in the case of viruses and virus‐like structures, part of these contaminants could be incorporated into the viral structure (Pato et al, 2014; Sviben, Forcic, Ivancic‐Jelecki, Halassy, & Brgles, 2017). The presence of host‐cell nucleic acids such as RNA or DNA is of general concern in the field of VLP.…”

Section: Resultsmentioning

confidence: 99%

Quantification of the HIV‐1 virus‐like particle production process by super‐resolution imaging: From VLP budding to nanoparticle analysis

Puente‐Massaguer

Cervera

Gòdia

2020

Biotech & Bioengineering

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Virus‐like particles (VLPs) offer great promise in the field of nanomedicine. Enveloped VLPs are a class of these nanoparticles and their production process occurs by a budding process, which is known to be the most critical step at intracellular level. In this study, we developed a novel imaging method based on super‐resolution fluorescence microscopy (SRFM) to assess the generation of VLPs in living cells. This methodology was applied to study the production of Gag VLPs in three animal cell platforms of reference: HEK 293‐transient gene expression (TGE), High Five‐baculovirus expression vector system (BEVS) and Sf9‐BEVS. Quantification of the number of VLP assembly sites per cell ranged from 500 to 3,000 in the different systems evaluated. Although the BEVS was superior in terms of Gag polyprotein expression, the HEK 293‐TGE platform was more efficient regarding the assembly of Gag as VLPs. This was translated into higher levels of non‐assembled Gag monomer in BEVS harvested supernatants. Furthermore, the presence of contaminating nanoparticles was evidenced in all three systems, specifically in High Five cells. The SRFM‐based method here developed was also successfully applied to measure the concentration of VLPs in crude supernatants. The lipid membrane of VLPs and the presence of nucleic acids alongside these nanoparticles could also be detected using common staining procedures. Overall, a complete picture of the VLP production process was achieved in these three production platforms. The robustness and sensitivity of this new approach broaden the applicability of SRFM toward the development of new detection, diagnosis and quantification methods based on confocal microscopy in living systems.

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Section: Resultsmentioning

confidence: 99%

Quantification of the HIV‐1 virus‐like particle production process by super‐resolution imaging: From VLP budding to nanoparticle analysis

Puente‐Massaguer

Cervera

Gòdia

2020

Biotech & Bioengineering

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“…TFF-based concentration/purification is used for the preparation of many virus products, and we therefore investigated its suitability for the production of oncolytic MV given the paucity of recent data on MV recovery and purification [18,21]. Indeed, the only articles dealing with TFF to reduce the volume of MV-containing suspensions were published in the 1970s and 1980s, and further purification was achieved by centrifugation [19,20].…”

Section: Discussionmentioning

confidence: 99%

“…In a previous report involving the purification of MV by hydrophobic interaction chromatography, the recovery of infectious MV increased when the R T/I of the feed was higher, and at the highest R T/I of 10 5 the authors achieved a recovery of~60%. [21]. Therefore, the outcome of TFF may be influenced by the behavior of the non-infective virus particles.…”

Section: Discussionmentioning

confidence: 99%

“…These studies focused on volume reduction rather than the characteristics of TFF, and provided data for virus recovery (62-100%) and in some cases the reduction of host cell protein levels [19,20]. After decades without further progress, the advent of oncolytic MV therapy has rekindled interest in the DSP steps, including the concentration/purification of MV produced in serum-free medium (SFM) [18,21]. MV purification is not trivial, and significant losses are often reported: up to 42% during microfiltration, 79-84% during centrifugal ultrafiltration, 83% during ion-exchange chromatography, and 40% during hydrophobic interaction chromatography [18,21].…”

Section: Introductionmentioning

confidence: 99%

“…After decades without further progress, the advent of oncolytic MV therapy has rekindled interest in the DSP steps, including the concentration/purification of MV produced in serum-free medium (SFM) [18,21]. MV purification is not trivial, and significant losses are often reported: up to 42% during microfiltration, 79-84% during centrifugal ultrafiltration, 83% during ion-exchange chromatography, and 40% during hydrophobic interaction chromatography [18,21]. The influence of the cell culture medium on DSP has not been investigated.…”

Section: Introductionmentioning

confidence: 99%

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Tangential Flow Filtration for the Concentration of Oncolytic Measles Virus: The Influence of Filter Properties and the Cell Culture Medium

et al. 2019

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The therapeutic use of oncolytic measles virus (MV) for cancer treatment requires >108 infectious MV particles per dose in a highly pure form. The concentration/purification of viruses is typically achieved by tangential flow filtration (TFF) but the efficiency of this process for the preparation of MV has not been tested in detail. We therefore investigated the influence of membrane material, feed composition, and pore size or molecular weight cut-off (MWCO) on the recovery of MV by TFF in concentration mode. We achieved the recovery of infectious MV particles using membranes with a MWCO ≤ 300 kDa regardless of the membrane material and whether or not serum was present in the feed. However, serum proteins in the medium affected membrane flux and promoted fouling. The severity of fouling was dependent on the membrane material, with the cellulose-based membrane showing the lowest susceptibility. We found that impurities such as proteins and host cell DNA were best depleted using membranes with a MWCO ≥ 300 kDa. We conclude that TFF in concentration mode is a robust unit operation to concentrate infectious MV particles while depleting impurities such as non-infectious MV particles, proteins, and host cell DNA.

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Vaccine process technology—A decade of progress

Buckland,

Sanyal,

Ranheim

et al. 2024

Biotech & Bioengineering

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In the past decade, new approaches to the discovery and development of vaccines have transformed the field. Advances during the COVID‐19 pandemic allowed the production of billions of vaccine doses per year using novel platforms such as messenger RNA and viral vectors. Improvements in the analytical toolbox, equipment, and bioprocess technology have made it possible to achieve both unprecedented speed in vaccine development and scale of vaccine manufacturing. Macromolecular structure‐function characterization technologies, combined with improved modeling and data analysis, enable quantitative evaluation of vaccine formulations at single‐particle resolution and guided design of vaccine drug substances and drug products. These advances play a major role in precise assessment of critical quality attributes of vaccines delivered by newer platforms. Innovations in label‐free and immunoassay technologies aid in the characterization of antigenic sites and the development of robust in vitro potency assays. These methods, along with molecular techniques such as next‐generation sequencing, will accelerate characterization and release of vaccines delivered by all platforms. Process analytical technologies for real‐time monitoring and optimization of process steps enable the implementation of quality‐by‐design principles and faster release of vaccine products. In the next decade, the field of vaccine discovery and development will continue to advance, bringing together new technologies, methods, and platforms to improve human health.

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Recovery of infective virus particles in ion-exchange and hydrophobic interaction monolith chromatography is influenced by particle charge and total-to-infective particle ratio

Cited by 30 publications

References 36 publications

Quantification of the HIV‐1 virus‐like particle production process by super‐resolution imaging: From VLP budding to nanoparticle analysis

Quantification of the HIV‐1 virus‐like particle production process by super‐resolution imaging: From VLP budding to nanoparticle analysis

Tangential Flow Filtration for the Concentration of Oncolytic Measles Virus: The Influence of Filter Properties and the Cell Culture Medium

Vaccine process technology—A decade of progress

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