Abstract
High throughput screening (HTS) is at the core of the drug discovery process, and so it is critical to design and implement HTS assays in a comprehensive fashion involving scientists from the disciplines of biology, chemistry, engineering, and informatics. This requires careful analysis of many variables, starting with the choice of assay target and ending with the discovery of lead compounds. At every step in this process, there are decisions to be made that can greatly impact the outcome of the HTS effort, to the point of making it a success or a failure. Although specific guidelines should be established to insure that the screening assay reaches an acceptable level of quality, many choices require pragmatism and the ability to compromise opposing forces.
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Abbreviations
- AAO:
-
Automated assay optimization
- AAS:
-
Atomic absorbance spectroscopy
- BRET:
-
Bioluminescence resonance energy transfer
- Bicine:
-
N,N-bis(2-Hydroxyethyl)glycine
- B max :
-
Maximum binding capacity
- BSA:
-
Bovine serum albumin
- CHAPS:
-
3-([3-Cholamidopropyl]dimethylammonio)-1-propanesulfonate
- CV:
-
Coefficient of variation
- DMSO:
-
Dimethyl sulfoxide
- DTT:
-
Dithiothreitol
- ECL:
-
Electrochemiluminescence
- EDTA:
-
Ethylenediamine-N,N,N′,N′-tetraacetic acid
- EFC:
-
Enzyme fragment complementation
- EGTA:
-
Ethylene glycol-bis(2-aminoethyl)-N,N,N′,N′-tetraacetic acid
- ELISA:
-
Enzyme-linked immunosorbent assay
- FCS:
-
Fluorescence correlation spectroscopy
- FIDA:
-
Fluorescence intensity distribution analysis
- FLINT:
-
Fluorescence intensity
- FRET:
-
Fluorescence resonance energy transfer
- FP:
-
Fluorescence polarization
- GPCR:
-
G-protein coupled receptor
- HTS:
-
High throughput screening
- K d :
-
Dissociation constant
- L:
-
Ligand
- M:
-
Mean
- NSB:
-
Non-specific binding
- OD:
-
Optical density unit
- PEI:
-
Polyethylene imine
- PMSF:
-
Phenylmethylsulfonyl fluoride
- RIA:
-
Radioimmunoassay
- S/B:
-
Signal to background ratio
- S/N:
-
Signal to noise ratio
- SD:
-
Standard deviation
- SW:
-
Signal window
- SPA:
-
Scintillation proximity assay
- TAPS:
-
N-tris(Hydroxymethyl)methyl-3-aminopropanesulfonic acid
- TR-FRET:
-
Time-resolved fluorescence resonance energy transfer
- V max :
-
Maximum velocity
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Acknowledgments
The authors are grateful to the many colleagues at GlaxoSmithKline that helped over the years to shape the screening process and to build the collective knowledge succinctly described in this introduction.
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Macarrón, R., Hertzberg, R.P. Design and Implementation of High Throughput Screening Assays. Mol Biotechnol 47, 270–285 (2011). https://doi.org/10.1007/s12033-010-9335-9
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DOI: https://doi.org/10.1007/s12033-010-9335-9