ArticleBiotechnology and bioengineering2019
Process monitoring of virus-like particle reassembly by diafiltration with UV/Vis spectroscopy and light scattering.
Article in Biotechnology and bioengineering, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- Downstream Purification Strategies for Virus-like Particles: A Systematic Review of Structure Preservation, Impurity Control, and Viral Safety.Microorganisms · 2026Review
- Ferritin-based nanoparticle vaccine protects neonatal piglets against porcine epidemic diarrhea virus challenge following immunization of pregnant sows.Veterinary research · 2025Article
- Raman-based PAT for multi-attribute monitoring during VLP recovery by dual-stage CFF: attribute-specific spectral preprocessing for model transfer.Frontiers in bioengineering and biotechnology · 2025Article
- Raman-based PAT for VLP precipitation: systematic data diversification and preprocessing pipeline identification.Frontiers in bioengineering and biotechnology · 2024Article
- Absolute Quantification of Hepatitis B Core Antigen (HBcAg) Virus-like Particles and Bound Nucleic Acids.Viruses · 2023Article
- Asymmetric parameter enhancement in the split-ring cavity array for virus-like particle sensing.Biomedical optics express · 2023Article
- Monitoring of ultra- and diafiltration processes by Kalman-filtered Raman measurements.Analytical and bioanalytical chemistry · 2023Article
- Size-selective downstream processing of virus particles and non-enveloped virus-like particles.Frontiers in bioengineering and biotechnology · 2023Review
- Article
- Review
- Fluorescence signatures of SARS-CoV-2 spike S1 proteins and a human ACE-2: excitation-emission maps and fluorescence lifetimes.Journal of biomedical optics · 2022Article
- Effects of Different Lengths of a Nucleic Acid Binding Region and Bound Nucleic Acids on the Phase Behavior and Purification Process of HBcAg Virus-Like Particles.Frontiers in bioengineering and biotechnology · 2022Article
- Soft Sensor Development for Real-Time Process Monitoring of Multidimensional Fractionation in Tubular Centrifuges.Nanomaterials (Basel, Switzerland) · 2021Article
- Multi-attribute PAT for UF/DF of Proteins-Monitoring Concentration, particle sizes, and Buffer Exchange.Analytical and bioanalytical chemistry · 2020Article
- A critical review of recent trends, and a future perspective of optical spectroscopy as PAT in biopharmaceutical downstream processing.Analytical and bioanalytical chemistry · 2020Review
- Integrated Process for Capture and Purification of Virus-Like Particles: Enhancing Process Performance by Cross-Flow Filtration.Frontiers in bioengineering and biotechnology · 2020Article
- Optimization of a Soft Ensemble Vote Classifier for the Prediction of Chimeric Virus-Like Particle Solubility and Other Biophysical Properties.Frontiers in bioengineering and biotechnology · 2020Article
- Ensembles of Hydrophobicity Scales as Potent Classifiers for Chimeric Virus-Like Particle Solubility - An Amino Acid Sequence-Based Machine Learning Approach.Frontiers in bioengineering and biotechnology · 2020Article
- Process monitoring of virus-like particle reassembly by diafiltration with UV/Vis spectroscopy and light scattering.Biotechnology and bioengineering · 2019Article
- Mechanistic deconvolution of BSA size variants by constrained Raman pseudo-Voigt hard modeling during anion-exchange chromatography.Biotechnology progressArticle
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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.
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