The Evolution of MALDI-TOF Mass Spectrometry toward Ultra-High-Throughput Screening: 1536-Well Format and Beyond
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 drama...
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| Published in | Journal of biomolecular screening Vol. 21; no. 2; pp. 176 - 186 |
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| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
01.02.2016
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| Subjects | |
| Online Access | Get full text |
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| 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. |
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| AbstractList | 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. |
| Author | Leveridge, Melanie Paape, Rainer Dunn, Adrian Pemberton, Michelle Haslam, Carl Marshall, Peter Hardy, Neil Fuetterer, Arne Hellicar, John Resemannand, Anja |
| Author_xml | – sequence: 1 givenname: Carl surname: Haslam fullname: Haslam, Carl – sequence: 2 givenname: John surname: Hellicar fullname: Hellicar, John – sequence: 3 givenname: Adrian surname: Dunn fullname: Dunn, Adrian – sequence: 4 givenname: Arne surname: Fuetterer fullname: Fuetterer, Arne – sequence: 5 givenname: Neil surname: Hardy fullname: Hardy, Neil – sequence: 6 givenname: Peter surname: Marshall fullname: Marshall, Peter – sequence: 7 givenname: Rainer surname: Paape fullname: Paape, Rainer – sequence: 8 givenname: Michelle surname: Pemberton fullname: Pemberton, Michelle – sequence: 9 givenname: Anja surname: Resemannand fullname: Resemannand, Anja – sequence: 10 givenname: Melanie surname: Leveridge fullname: Leveridge, Melanie |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/26428484$$D View this record in MEDLINE/PubMed |
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| SubjectTerms | Acetylcholinesterase - chemistry Chromatography, Liquid - methods High-Throughput Screening Assays - methods Histone Demethylases - chemistry Peptides - chemistry Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods |
| Title | The Evolution of MALDI-TOF Mass Spectrometry toward Ultra-High-Throughput Screening: 1536-Well Format and Beyond |
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