Biophysical Characterization and Conformational Stability of Ebola and Marburg Virus-Like Particles

Hu, Lei; Trefethen, Jared M.; Zeng, Yuhong; Yee, Luisa; Ohtake, Satoshi; Lechuga‐Ballesteros, David; Warfield, Kelly L.; Aman, M. Javad; Shulenin, Sergey; Unfer, Robert C.; Enterlein, Sven; Truong‐Le, Vu; Volkin, David B.; Joshi, Sangeeta B.; Middaugh, C. Russell

doi:10.1002/jps.22724

Cited by 25 publications

(15 citation statements)

References 47 publications

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“…The decrease in light scattering intensity may also reflect a change in the differential refractive index (d n /d c ). This decrease in light scattering intensity has been previously observed in human adenovirus type 4, measles virus, and filovirus virus‐like particles (VLPs) . In these studies, as well as the current study, there is no direct evidence to support or disprove the precise mechanism.…”

Section: Discussioncontrasting

confidence: 60%

Characterization of an Oncolytic Herpes Simplex Virus Drug Candidate

Kumru

Joshi

Thapa

et al. 2015

Journal of Pharmaceutical Sciences

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Section: Discussioncontrasting

confidence: 60%

Characterization of an Oncolytic Herpes Simplex Virus Drug Candidate

Kumru

Joshi

Thapa

et al. 2015

Journal of Pharmaceutical Sciences

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“…Particle size measurements provide information on the assembly progress of the capsid proteins into VLPs. Previous publications have also reported effects of the VLP tertiary structure on ultraviolet and visible (UV/Vis) and fluorescence absorption spectra (Ausar, Foubert, Hudson, Vedvick, & Middaugh, 2006;Fang et al, 2016;Hanslip, Zaccai, Middelberg, & Falconer, 2006;Hu et al, 2011;Rajendar et al, 2013). Following a systematic approach to process monitoring, a combination of PAT sensors should be chosen which allows monitoring protein concentration, protein tertiary structure, and protein size.…”

mentioning

confidence: 99%

Process monitoring of virus‐like particle reassembly by diafiltration with UV/Vis spectroscopy and light scattering

Rüdt

Vormittag

Hillebrandt

et al. 2019

Biotech & Bioengineering

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Virus‐like particles (VLPs) have shown great potential as biopharmaceuticals in the market and in clinics. Nonenveloped, in vivo assembled VLPs are typically disassembled and reassembled in vitro to improve particle stability, homogeneity, and immunogenicity. At the industrial scale, cross‐flow filtration (CFF) is the method of choice for performing reassembly by diafiltration. Here, we developed an experimental CFF setup with an on‐line measurement loop for the implementation of process analytical technology (PAT). The measurement loop included an ultraviolet and visible (UV/Vis) spectrometer as well as a light scattering photometer. These sensors allowed for monitoring protein concentration, protein tertiary structure, and protein quaternary structure. The experimental setup was tested with three Hepatitis B core Antigen (HBcAg) variants. With each variant, three reassembly processes were performed at different transmembrane pressures (TMPs). While light scattering provided information on the assembly progress, UV/Vis allowed for monitoring the protein concentration and the rate of VLP assembly based on the microenvironment of Tyrosine‐132. VLP formation was verified by off‐line dynamic light scattering (DLS) and transmission electron microscopy (TEM). Furthermore, the experimental results provided evidence of aggregate‐related assembly inhibition and showed that off‐line size‐exclusion chromatography does not provide a complete picture of the particle content. Finally, a Partial‐Least Squares (PLS) model was calibrated to predict VLP concentrations in the process solution. Q 2 values of 0.947–0.984 were reached for the three HBcAg variants. In summary, the proposed experimental setup provides a powerful platform for developing and monitoring VLP reassembly steps by CFF.

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“…VLP biophysical characterization with quantitative outputs is accomplished through dynamic light scattering (DLS; Hu et al, 2011;Kissmann et al, 2011), disc centrifugation particle size analysis (Deschuyteneer et al, 2010), electrospray differential mobility analysis (ES-DMA; Pease et al, 2009), and asymmetric flow field-flow fractionation coupled with multiple-angle light scattering (AF4-MALS; Chuan et al, 2008;Citkowicz et al, 2008;Lang et al, 2009Lang et al, , 2010Mohr et al, 2013;Pease et al, 2009). DLS is the simplest to perform and can accurately and quickly determine the particle size of a monodispersed sample.…”

Section: Vlp Characterizationmentioning

confidence: 99%

Bioengineering virus‐like particles as vaccines

Lua

Connors

Sainsbury

et al. 2013

Biotech & Bioengineering

314

268

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Virus-like particle (VLP) technology seeks to harness the optimally tuned immunostimulatory properties of natural viruses while omitting the infectious trait. VLPs that assemble from a single protein have been shown to be safe and highly efficacious in humans, and highly profitable. VLPs emerging from basic research possess varying levels of complexity and comprise single or multiple proteins, with or without a lipid membrane. Complex VLP assembly is traditionally orchestrated within cells using black-box approaches, which are appropriate when knowledge and control over assembly are limited. Recovery challenges including those of adherent and intracellular contaminants must then be addressed. Recent commercial VLPs variously incorporate steps that include VLP in vitro assembly to address these problems robustly, but at the expense of process complexity. Increasing research activity and translation opportunity necessitate bioengineering advances and new bioprocessing modalities for efficient and cost-effective production of VLPs. Emerging approaches are necessarily multi-scale and multi-disciplinary, encompassing diverse fields from computational design of molecules to new macro-scale purification materials. In this review, we highlight historical and emerging VLP vaccine approaches. We overview approaches that seek to specifically engineer a desirable immune response through modular VLP design, and those that seek to improve bioprocess efficiency through inhibition of intracellular assembly to allow optimal use of existing purification technologies prior to cell-free VLP assembly. Greater understanding of VLP assembly and increased interdisciplinary activity will see enormous progress in VLP technology over the coming decade, driven by clear translational opportunity.

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Biophysical Characterization and Conformational Stability of Ebola and Marburg Virus-Like Particles

Cited by 25 publications

References 47 publications

Characterization of an Oncolytic Herpes Simplex Virus Drug Candidate

Characterization of an Oncolytic Herpes Simplex Virus Drug Candidate

Process monitoring of virus‐like particle reassembly by diafiltration with UV/Vis spectroscopy and light scattering

Bioengineering virus‐like particles as vaccines

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