Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Analysis of baculovirus aggregates using flow cytometry.

Hasnaa Jorio1, Rosa Tran, Jamal Meghrous

  • 1Animal Cell Technology Group, Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Ave., Montreal, Que., Canada H1K 4S9.

Journal of Virological Methods
|December 21, 2005
PubMed
Summary

Flow cytometry can detect baculovirus aggregates, which form during storage or under specific pH conditions. This method uses fluorescence intensity to indicate aggregate presence and size, improving viral stock quality control.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Impact of Lectin biotinylation for surface plasmon resonance and enzyme-linked Lectin assays for protein glycosylation.

Analytical biochemistry·2024
Same author

Impact of Recombinant VSV-HIV Prime, DNA-Boost Vaccine Candidates on Immunogenicity and Viremia on SHIV-Infected Rhesus Macaques.

Vaccines·2024
Same author

Antibody-independent surface plasmon resonance assays for influenza vaccine quality control.

Applied microbiology and biotechnology·2024
Same author

Production of recombinant vesicular stomatitis virus-based vectors by tangential flow depth filtration.

Applied microbiology and biotechnology·2024
Same author

Transient Expression in HEK-293 Cells in Suspension Culture as a Rapid and Powerful Tool: SARS-CoV-2 N and Chimeric SARS-CoV-2N-CD154 Proteins as a Case Study.

Biomedicines·2023
Same author

Membrane Chromatography-Based Downstream Processing for Cell-Culture Produced Influenza Vaccines.

Vaccines·2022

Area of Science:

  • Virology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Baculovirus particle aggregation poses challenges in stock processing, storage, and accurate particle counting.
  • A previously established flow cytometry method for baculovirus concentration measurement was adapted for aggregate detection.

Purpose of the Study:

  • To extend the application of flow cytometry for detecting baculovirus aggregates.
  • To investigate the characteristics of baculovirus aggregates using flow cytometry.

Main Methods:

  • Baculovirus stocks were analyzed using flow cytometry after staining with SYBR Green.
  • Aggregation was induced by storage at 4°C or by altering pH to 5.3.
  • Treatment with endonuclease (Benzonase) and filtration experiments were performed to characterize the aggregated particles.

Related Experiment Videos

Main Results:

  • Fresh baculovirus stocks showed a unimodal distribution, while stored stocks exhibited a bimodal distribution indicating aggregation.
  • The aggregated population displayed high fluorescence intensity and consisted of particles larger than single baculoviruses.
  • Induced aggregation at pH 5.3 also resulted in a bimodal distribution, confirming flow cytometry's sensitivity to aggregation.

Conclusions:

  • Flow cytometry is a viable method for detecting baculovirus aggregates.
  • High fluorescence intensity populations in flow cytometry analysis correlate with baculovirus aggregates.
  • Mean green fluorescence intensity can serve as an indicator of mean aggregate size, aiding in quality control.