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Related Concept Videos

Upstream Processing01:27

Upstream Processing

Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
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Vaccine production involves a sequence of upstream and downstream processes to generate a safe and effective immunological product. It begins with cultivating microorganisms, such as viruses or bacteria, to obtain antigenic material. For viral vaccines, mammalian host cells are grown in bioreactors and subsequently infected with the target virus. The virus replicates within the host cells, which are lysed to release viral particles. This lysate is then clarified through filtration or...

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Related Experiment Video

Updated: Jun 16, 2026

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A Bioreactor-Based Yellow Fever Virus-like Particle Production Process with Integrated Process Analytical Technology

Gregor Dekevic1, Tobias Tertel2, Lars Tasto1

  • 1Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, 35390 Giessen, Germany.

Viruses
|October 28, 2023
PubMed
Summary

Developing a Yellow Fever vaccine-like particle (YF-VLP) bioreactor process offers a new passive immunization strategy. This method optimizes plasmid DNA production and VLP yield for potential use against neglected tropical diseases.

Keywords:
HEK 293T cellsIFCMProcess Analytical Technology (PAT)design of experimentslPEIstirred-tank bioreactor (STR)

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Area of Science:

  • Biotechnology
  • Virology
  • Bioprocess Engineering

Background:

  • Yellow Fever (YF) is a severe viral disease with no rapid treatment options for infected individuals.
  • Passive immunization using YF-virus-like particles (YF-VLPs) presents a potential therapeutic avenue.
  • Existing YF prevention relies solely on vaccination, highlighting the need for alternative interventions.

Purpose of the Study:

  • To establish a bioreactor-based production process for Yellow Fever virus-like particles (YF-VLPs).
  • To optimize plasmid DNA (pDNA) production for enhanced VLP yield.
  • To integrate Process Analytical Technology (PAT) for real-time process monitoring and control.

Main Methods:

  • Utilized design of experiments to optimize pDNA production, identifying key media components influencing yield.
  • Adjusted HEK cell density, polyplex formation duration, and medium exchange strategies to improve transfection efficiency.
  • Implemented a stirred-tank bioreactor with integrated dielectric spectroscopy for real-time monitoring of cell growth, polyplex uptake, and harvest timing.

Main Results:

  • Achieved a plasmid DNA yield of 11 mg/L, with glucose, NaCl, yeast extract, and phosphate buffer significantly impacting specific pDNA yield.
  • Demonstrated increased transfection efficiencies through optimized bioreactor conditions and identified Pluronic F-68 as neutral and anti-clumping agents as detrimental.
  • Successfully confirmed YF-VLP presence and integrity using Western blot, imaging flow cytometry, and transmission electron microscopy.

Conclusions:

  • Established a robust bioreactor process for producing Yellow Fever virus-like particles (YF-VLPs).
  • The developed YF-VLP production process can be adapted as a platform for generating VLPs against other neglected tropical diseases.
  • This work provides a foundation for developing passive immunization strategies against viral infections.