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A simple method for obtaining highly viable virus from culture supernatant
1University of Texas, M.D. Anderson Cancer Center, Science Park-Research Division, Smithville 78957.
Journal of Virological Methods
|March 1, 1994
Summary
Ultrafiltration offers a faster, more effective method for concentrating viruses than traditional density-gradient techniques. This single-step approach preserves viral infectivity, crucial for applications like virus-cell binding studies.
Area of Science:
- Virology
- Biotechnology
- Molecular Biology
Background:
- Traditional virus purification methods like density-gradient centrifugation are time-consuming and result in low recovery of viable viruses.
- These limitations hinder downstream applications requiring high concentrations of infectious viral particles.
Purpose of the Study:
- To evaluate ultrafiltration as a rapid, single-step concentration technique for Moloney murine leukemia virus (ts1).
- To compare the infectivity recovery of viruses concentrated by ultrafiltration versus traditional methods.
Main Methods:
- Concentration of ts1 virus from culture supernatants using a single-step ultrafiltration unit with a 100,000 mol wt cut-off.
- Comparison of viral recovery and infectivity with viruses purified by traditional sucrose density gradient methods.
Main Results:
- Ultrafiltration achieved approximately 30-fold virus concentration without loss of infectivity.
- Traditional sucrose density gradient purification resulted in significant loss of viral infectivity.
Conclusions:
- Ultrafiltration is a superior method for concentrating viruses, offering speed, simplicity, and high infectivity recovery.
- This technique is broadly applicable for concentrating various viruses for research purposes, including virus-cell binding assays.