Experimental design and statistical methods for improved hit detection in high-throughput screening.
Identification of active compounds in high-throughput screening (HTS) contexts can be substantially improved by applying classical experimental design and statistical inference principles to all phases of HTS studies. The authors present both experimental and simulated data to illustrate how true-positive rates can be maximized without increasing false-positive rates by the following analytical process. First, the use of robust data preprocessing methods reduces unwanted variation by removing row, column, and plate biases. Second, replicate measurements allow estimation of the magnitude of the remaining random error and the use of formal statistical models to benchmark putative hits relative to what is expected by chance. Receiver Operating Characteristic (ROC) analyses revealed superior power for data preprocessed by a trimmed-mean polish method combined with the RVM t-test, particularly for small- to moderate-sized biological hits.
Malo
Nathalie
N
McGill University and Genome Quebec Innovation Centre, Montreal, Quebec, Canada.
Hanley
James A
JA
Carlile
Graeme
G
Liu
Jing
J
Pelletier
Jerry
J
Thomas
David
D
Nadon
Robert
R
D000818:Animals; D002474:Cell-Free System / Q000187:drug effects; D003198:Computer Simulation; D004353:Drug Evaluation, Preclinical / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data; D005189:False Positive Reactions; D005455:Fluorescent Antibody Technique / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data; D057166:High-Throughput Screening Assays / Q000379:methods / Q000592:standards* / Q000706:statistics & numerical data*; D049409:Luciferases, Firefly / Q000032:analysis / Q000378:metabolism; D049410:Luciferases, Renilla / Q000032:analysis / Q000378:metabolism; D015233:Models, Statistical; D014176:Protein Biosynthesis / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000302:isolation & purification / Q000494:pharmacology; D012372:ROC Curve; D011897:Random Allocation; D012107:Research Design
D011500:Protein Synthesis Inhibitors; D049410:Luciferases, Renilla; D049409:Luciferases, Firefly