Hello,
During the development of ELISAs, I always debate with myself for if I should put CV% for zero standard into the assay performance evaluation.
For processing data, I usually subtract the absorbance of zero-standard from the sample and non-zero standards, than calculate the CV%. But after this operation, the CV% of zero-standard has great opportunity higher than 25%. For example, in a triplicate case, the zero-standard ODs are 0.001, 0.002, and -0.001. The CV% is 229%, but the ODs actually make sense for itself. The CV% fail only because the absolute absorbance is too low.
This issue could be avoid if we don't include the zero-standard into standard curve, then we don't have to evaluate the CV% of it.
Now question is, is it popular that people don't include the zero-standard into standard curve? Based on what I see, it is quite amount assay include zero-standard, but the other doesn't.
What do you do when the CV% of your zero-standard is higher then 20% (after all standards subtract the zero-standard's absorbance)?
Thank you very much for any input!
For more detail on the use of CV% for the ELISA performance evaluation see the published article given below:
Optimization, Validation and Standardization of ELISA
By Rajna Minic and Irena Zivkovic. DOI: 10.5772/intechopen.94338
Chapter Optimization, Validation and Standardization of ELISA
Hope this article may be useful.
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