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The Interferon Protection Assay
29 August 2011
Why:
The drug company our lab works for wants to test their antibodies conjugated to IFNs for antiviral activity. My task is to understand how much antibody-IFN is needed to "cure" an organism from a viral infection (specifically viruses which causes phosphatidylserine to be expressed on the outside of the cellular membrane.)
What Happened:
After the EMCV (Encephalomyocarditis virus) kill curve (using L929 cells) I was able to attain the specific dilution (for the inoculum) which kills half the amount of cells plated. Now its on to the interferon protection assay. Using the same cell strain at the same density (within a 96-welled plate); I plated 6 plates with 100uls of cells/media and let incubate at 37C in 5%C02.
On the agenda for tomorrow:
After realizing where these experiments are leading too; I discovered that, EMCV is not the main virus understudy. The drug company is not concerned with EMCV. These preliminary experiments are being done only to attain data which will be used for a completely different virus and cell type. Is this a correct practice ?
The drug company our lab works for wants to test their antibodies conjugated to IFNs for antiviral activity. My task is to understand how much antibody-IFN is needed to "cure" an organism from a viral infection (specifically viruses which causes phosphatidylserine to be expressed on the outside of the cellular membrane.)
What Happened:
After the EMCV (Encephalomyocarditis virus) kill curve (using L929 cells) I was able to attain the specific dilution (for the inoculum) which kills half the amount of cells plated. Now its on to the interferon protection assay. Using the same cell strain at the same density (within a 96-welled plate); I plated 6 plates with 100uls of cells/media and let incubate at 37C in 5%C02.
On the agenda for tomorrow:
- Inspect the plates for cellular health; proceed if all is well.
- Need to filter some more MILLIPORE water
- plate some more cells
- toss out the T25 flask containing 3T6 cells
- read up on phosphatidylserine
After realizing where these experiments are leading too; I discovered that, EMCV is not the main virus understudy. The drug company is not concerned with EMCV. These preliminary experiments are being done only to attain data which will be used for a completely different virus and cell type. Is this a correct practice ?
