High-resolution dose-response screening using droplet-based microfluidics.
A critical early step in drug discovery is the screening of a chemical library. Typically, promising compounds are identified in a primary screen and then more fully characterized in a dose-response analysis with 7-10 data points per compound. Here, we describe a robust microfluidic approach that increases the number of data points to approximately 10,000 per compound. The system exploits Taylor-Aris dispersion to create concentration gradients, which are then segmented into picoliter microreactors by droplet-based microfluidics. The large number of data points results in IC(50) values that are highly precise (± 2.40% at 95% confidence) and highly reproducible (CV = 2.45%, n = 16). In addition, the high resolution of the data reveals complex dose-response relationships unambiguously. We used this system to screen a chemical library of 704 compounds against protein tyrosine phosphatase 1B, a diabetes, obesity, and cancer target. We identified a number of novel inhibitors, the most potent being sodium cefsulodine, which has an IC(50) of 27 ± 0.83 μM.
Institut de Science et d'Ingénierie Supramoléculaires, Université de Strasbourg, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7006, 8 allée Gaspard Monge, F-67083 Strasbourg Cedex, France.
Oliver J
OJ
Miller
Abdeslam
A
El Harrak
Thomas
T
Mangeat
Jean-Christophe
JC
Baret
Lucas
L
Frenz
Bachir
B
El Debs
Estelle
E
Mayot
Michael L
ML
Samuels
Eamonn K
EK
Rooney
Pierre
P
Dieu
Martin
M
Galvan
Darren R
DR
Link
Andrew D
AD
Griffiths
D002441:Cefsulodin / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005453:Fluorescence; D020128:Inhibitory Concentration 50; D044085:Microfluidics / Q000379:methods*; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1 / Q000037:antagonists & inhibitors; D018401:Sample Size; D054852:Small Molecule Libraries; D001616:beta-Galactosidase / Q000037:antagonists & inhibitors
D054852:Small Molecule Libraries; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D001616:beta-Galactosidase; D002441:Cefsulodin