could someone help me to understand the following sentence better? It's from the book "A Practical Guide to Assay Development and High-Throughput Screening in Drug Discovery". And it's about fluorophors used in FRET-assays, for screening of protease inhibitors.

"In contrast to many other FRET dye pairs, both EDANS & DABCYL and Mca/Dnp generate a dynamic range between cleaved an uncleaved substrate, sufficent for the application of such protease assays in HTS and high-throughput compound profiling under initial conditions. "

I'm confused about the last part of the quote "under initial conditions". What does that mean? the question summarized "What is the dynamic range under initial conditions? "


1 Answer 1


I think that they mean initial profiling as the profiling/screening is the subject of the clause.

In this case it would mean that FRET with these compounds has enough difference between cleaved and uncleaved that you can use this difference to tell if it has inhibited or promoted a reaction between your target and its substrate or not in a quick/high-throughput assay for mass screening of potential inhibitors/promoters.

  • $\begingroup$ I've read that its common to do HTS screening first with an easy and cheap assay. Afterwards more complex assays follow, which sort out more substances. So "initial" in this case would mean, something like: "the first assay to screen for substances?" $\endgroup$
    – Mourinho_1
    Oct 15, 2021 at 10:05
  • 1
    $\begingroup$ @Mourinho_1 that's how I read it. $\endgroup$
    – bob1
    Oct 15, 2021 at 11:37
  • $\begingroup$ thanks a lot. I did not consider that it could mean something like this. I would say that this sounds plausible $\endgroup$
    – Mourinho_1
    Oct 15, 2021 at 11:47

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