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Evaluating Novel Quantification Methods for Infectious Baculoviruses.

Keven Lothert1, Elena Bagrin1, Michael W Wolff1

  • 1Institute of Bioprocess Engineering and Pharmaceutical Technology, Department Life Science Engineering, University of Applied Sciences Mittelhessen (THM), 35390 Giessen, Germany.

Viruses
|April 28, 2023
PubMed
Summary
This summary is machine-generated.

Accurate virus quantification is crucial for vaccine production. Flow cytometry, particularly surface protein staining, offers a rapid and feasible method for assessing infectious virus titers, outperforming traditional assays.

Keywords:
flow cytometrygp64 proteininfectivity titernative baculovirusprocess monitoringquantitative polymerase chain reactionviral RNAvirus quantification

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Area of Science:

  • Biotechnology
  • Virology
  • Process Analytical Technology

Background:

  • Accurate quantification of infectious virus titers is essential for viral vector and vaccine manufacturing.
  • Current methods like endpoint dilution assays are time-consuming and not suitable for real-time process monitoring.
  • Flow cytometry and quantitative polymerase chain reaction (qPCR) are emerging as faster alternatives.

Purpose of the Study:

  • To compare different methods for assessing infectious virus titers.
  • To evaluate flow cytometry techniques and qPCR for rapid viral quantification.
  • To investigate the potential of viral nucleic acid labeling for infectivity assessment.

Main Methods:

  • Quantification of viral nucleic acids in infected cells using qPCR.
  • Flow cytometry analysis based on post-infection fluorophore expression.
  • Flow cytometry analysis using fluorescent antibodies to label viral surface proteins.
  • Exploration of viral (messenger)RNA labeling in infected cells.

Main Results:

  • qPCR-based infectivity assessment requires significant method optimization.
  • Flow cytometry using viral surface protein staining is a fast and practical approach for enveloped viruses.
  • Labeling viral (messenger)RNA in infected cells shows promise but needs further investigation.

Conclusions:

  • Flow cytometry, especially surface protein staining, provides a rapid and effective method for quantifying infectious virus titers.
  • While qPCR can be used, it demands complex optimization for accurate infectivity assessment.
  • Viral (messenger)RNA labeling presents a potential future avenue for infectivity analysis.