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Design and Implementation of High Throughput Screening Assays

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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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Authors and Affiliations

  1. Molecular Discovery Research, GlaxoSmithKline, 1250 S Collegeville Road, Collegeville, PA, 19426, USA

    Ricardo Macarrón & Robert P. Hertzberg

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  1. Ricardo Macarrón
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  2. Robert P. Hertzberg
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Correspondence to Ricardo Macarrón.

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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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