title: High-throughput screening assay for sphingosine kinase inhibitors in whole blood using RapidFire® mass spectrometry.
abstract: To facilitate discovery of compounds modulating sphingosine-1-phosphate (S1P) signaling, the authors used high-throughput mass spectrometry technology to measure S1P formation in human whole blood. Since blood contains endogenous sphingosine (SPH) and S1P, mass spectrometry was chosen to detect the conversion of an exogenously added 17-carbon-long variant of sphingosine, C17SPH, into C17S1P. The authors developed procedures to achieve homogeneous mixing of whole blood in 384-well plates and for a method requiring minimal manipulations to extract S1P from blood in 96- and 384-well plates prior to analyses using the RapidFire(®) mass spectrometry system.
authors.affiliation: Pfizer Global R&D, St. Louis, MO 63017, USA. maureen.highkin@gmail.com
authors.forename: Maureen K
authors.initials: MK
authors.lastname: Highkin
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Yates
authors.forename: Olga V
authors.initials: OV
authors.lastname: Nemirovskiy
authors.forename: William A
authors.initials: WA
authors.lastname: Lamarr
authors.forename: Grace E
authors.initials: GE
authors.lastname: Munie
authors.forename: John W
authors.initials: JW
authors.lastname: Rains
authors.forename: Jaime L
authors.initials: JL
authors.lastname: Masferrer
authors.forename: Marek M
authors.initials: MM
authors.lastname: Nagiec
mesh_terms: D000627:Aminophenols / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000097:blood*; D057166:High-Throughput Screening Assays; D006801:Humans; D007700:Kinetics; D008246:Lysophospholipids / Q000378:metabolism; D013058:Mass Spectrometry; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism / Q000494:pharmacology; D000099157:Sphingosine Kinase
chemical_list: D000627:Aminophenols; D004791:Enzyme Inhibitors; D008246:Lysophospholipids; D017853:Phosphotransferases (Alcohol Group Acceptor); D013110:Sphingosine; D013844:Thiazoles; D000099157:Sphingosine Kinase; C525025:4-(4-(4-chloro-phenyl)thiazol-2-ylamino)phenol; C060506:sphingosine 1-phosphate