title: The Evolution of MALDI-TOF Mass Spectrometry toward Ultra-High-Throughput Screening: 1536-Well Format and Beyond.
abstract: Mass spectrometry (MS) offers a label-free, direct-detection method, in contrast to fluorescent or colorimetric methodologies. Over recent years, solid-phase extraction-based techniques, such as the Agilent RapidFire system, have emerged that are capable of analyzing samples in <10 s. While dramatically faster than liquid chromatography-coupled MS, an analysis time of 8-10 s is still considered relatively slow for full-diversity high-throughput screening (HTS). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) offers an alternative for high-throughput MS detection. However, sample preparation and deposition onto the MALDI target, as well as interference from matrix ions, have been considered limitations for the use of MALDI for screening assays. Here we describe the development and validation of assays for both small-molecule and peptide analytes using MALDI-TOF coupled with nanoliter liquid handling. Using the JMJD2c histone demethylase and acetylcholinesterase as model systems, we have generated robust data in a 1536 format and also increased sample deposition to 6144 samples per target. Using these methods, we demonstrate that this technology can deliver fast sample analysis time with low sample volume, and data comparable to that of current RapidFire assays.
authors.affiliation: Department of Biological Sciences, GlaxoSmithKline, Stevenage, UK.
authors.forename: Carl
authors.initials: C
authors.lastname: Haslam
authors.affiliation: Department of Biological Sciences, GlaxoSmithKline, Stevenage, UK.
authors.forename: John
authors.initials: J
authors.lastname: Hellicar
authors.affiliation: Department of Chemical Sciences, GlaxoSmithKline, Stevenage, UK.
authors.forename: Adrian
authors.initials: A
authors.lastname: Dunn
authors.affiliation: Bruker Daltonik GmbH, Bremen, Germany.
authors.forename: Arne
authors.initials: A
authors.lastname: Fuetterer
authors.affiliation: Department of Sample Management and Automation, GlaxoSmithKline, Stevenage, UK.
authors.forename: Neil
authors.initials: N
authors.lastname: Hardy
authors.affiliation: Department of Chemical Sciences, GlaxoSmithKline, Stevenage, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Marshall
authors.affiliation: Bruker Daltonik GmbH, Bremen, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Paape
authors.affiliation: Department of Biological Sciences, GlaxoSmithKline, Stevenage, UK.
authors.forename: Michelle
authors.initials: M
authors.lastname: Pemberton
authors.affiliation: Bruker Daltonik GmbH, Bremen, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Resemannand
authors.affiliation: Department of Biological Sciences, GlaxoSmithKline, Stevenage, UK melanie.v.leveridge@gsk.com.
authors.forename: Melanie
authors.initials: M
authors.lastname: Leveridge
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D056466:Histone Demethylases / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D010455:Peptides; D056466:Histone Demethylases; D000110:Acetylcholinesterase
keywords: MALDI-TOF; RapidFire; high-throughput screening; mass spectrometry