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Assessing HTS performance using BioAssay Ontology: screening and analysis of a
bacterial phospho-N-acetylmuramoyl-pentapeptide translocase campaign
#MMPMID25415593
Moberg A
; Zander Balderud L
; Hansson E
; Boyd H
Assay Drug Dev Technol
2014[Nov]; 12
(9-10
): 506-13
PMID25415593
show ga
With the public availability of biochemical assays and screening data constantly
increasing, new applications for data mining and method analysis are evolving in
parallel. One example is BioAssay Ontology (BAO) for systematic classification of
assays based on screening setup and metadata annotations. In this article we
report a high-throughput screening (HTS) against
phospho-N-acetylmuramoyl-pentapeptide translocase (MraY), an attractive
antibacterial drug target involved in peptidoglycan synthesis. The screen
resulted in novel chemistry identification using a fluorescence resonance energy
transfer assay. To address a subset of the false positive hits, a frequent hitter
analysis was performed using an approach in which MraY hits were compared with
hits from similar assays, previously used for HTS. The MraY assay was annotated
according to BAO and three internal reference assays, using a similar assay
design and detection technology, were identified. Analyzing the assays
retrospectively, it was clear that both MraY and the three reference assays all
showed a high false positive rate in the primary HTS assays. In the case of MraY,
false positives were efficiently identified by applying a method to correct for
compound interference at the hit-confirmation stage. Frequent hitter analysis
based on the three reference assays with similar assay method identified
additional false actives in the primary MraY assay as frequent hitters. This
article demonstrates how assays annotated using BAO terms can be used to identify
closely related reference assays, and that analysis based on these assays clearly
can provide useful data to influence assay design, technology, and screening
strategy.
|Biological Assay/*methods/standards
[MESH]
|Escherichia coli Proteins/*analysis
[MESH]
|Fluorescence Resonance Energy Transfer/methods/standards
[MESH]