Mar 18, 2025
Generating Antiviral & Cytotoxicity Curves
- 1Icahn School of Medicine at Mount Sinai;
- 2ASAP Discovery Consortium
Protocol Citation: Briana L McGovern 2025. Generating Antiviral & Cytotoxicity Curves. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvw12kzlmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 07, 2024
Last Modified: March 18, 2025
Protocol Integer ID: 104938
Keywords: prism, antiviral, drug discovery, cytotoxicity, dose response curve,
Funders Acknowledgements:
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
Grant ID: U19AI171399
Abstract
This describes generating Antiviral & Cytotoxicity Curves
Analyzing the Cytotoxicity Plates
Analyzing the Cytotoxicity Plates
After you scan the plates and export the data to excel you will have an output that looks like this:
If you used the MTS kit you will only have one
If you used the Roche MTT kit you will have two readings
MTS plate data
As a reminder, this is the typical layout of the plates:
Typical plate layout.
If you used the MTT kit you will first subtract the values from the two read outs:
First, take calculate the average value and standard deviation of columns 1 and 11. These are the DMSO controls.
DMSO controls columns 1 and 11 highlighted.
Next, you will take the average of the three replicates for each dilution of the compounds.
The triplicates of the first dilution of drug 1 on this plate.
The triplicates of the second dilution of drug 1 on this plate.
Next, you will divide the averages and standard deviations of each dilution by those of the DMSO controls and turn it into a percentage.
For simplicity in excel you can combine steps 3 and 4 into one formula:
=(average(A2-A4) / average(columns 1 & 11)) *100
=(standard deviation(A2-A4) / standard deviation(columns 1 & 11)) *100
Output
Plug these values into a Prism file for the cell viability section.
Analyzing the Antiviral Plates
Analyzing the Antiviral Plates
After staining and reading the plates you will export the data to an excel sheet. The data set that we want to further analyze is the ratio of infected to uninfected cells.
It should look like this:
Antiviral plate data output. Ratio of infected to uninfected cells.
Again, take the average of the infected DMSO controls. Columns 1 and 11.
infected DMSO controls highlighted.
Next, take the average of the uninfected control (column 12)
uninfected control highlighted
then, calculate the following using excel.
[((value of experimental well) - (average uninfected)) / (average infected DMSO controls) ]*100
So, from the above example data it will be:
[(2A- value from step 8) / (value from step 7)] *100
[(16.832-0.240375)/(24.684063)] = 67.21594
The data will look like this:
Analysis of data from this example experiment.
From the output of step 9, calculate the average and standard deviation between the triplicates
Left: Data output from step 9
Right: Average and standard deviation of the triplicates from step 9.
Plug these values into a Prism file for the % infection section.
Prism Output
Prism Output
This is what the final Prism output should look like.