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High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
ELECTROPHORETIC STUDIES ON PR8 INFLUENZA VIRUS
G L Miller1, M A Lauffer, W M Stanley
1Department of Animal and Plant Pathology of The Rockefeller Institute for Medical Research, Princeton, New Jersey.
The Journal of Experimental Medicine
|October 30, 2009
Summary
This study purified the PR8 influenza virus, separating the active virus from embryonic impurities using fractional centrifugation. The purified virus fraction was homogeneous and had an isoelectric point of pH 5.3.
Area of Science:
- Virology
- Biochemistry
Background:
- Crude preparations of PR8 influenza virus contain multiple components.
- Previous research identified an acidic impurity in uninfected embryos.
Purpose of the Study:
- To characterize and purify the active component of PR8 influenza virus.
- To separate the virus from embryonic impurities.
Main Methods:
- High-speed centrifugation to obtain crude virus preparations.
- Electrophoresis, centrifuge, and activity tests for component identification.
- Repeated fractional centrifugation for virus separation.
- Analytical ultracentrifugation and Tiselius electrophoresis for homogeneity assessment.
- Micro-electrophoresis to determine the isoelectric point.
Main Results:
- Crude PR8 influenza virus preparations contained two electrophoretically distinct components.
- An impurity (10-20%) was identified as a high molecular weight acidic substance from embryos.
- The active virus fraction (80-90%) was successfully separated from the impurity.
- The purified virus fraction appeared homogeneous and had an isoelectric point of pH 5.3.
Conclusions:
- PR8 influenza virus can be purified from embryonic impurities using fractional centrifugation.
- The purified virus is a distinct entity with a measurable isoelectric point.
- This purification method yields a homogeneous viral preparation suitable for further study.
