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Automated nano-electrospray mass spectrometry for protein-ligand screening by noncovalent interaction applied to human H-FABP and A-FABP

Benkestock, K ; van Pelt, C K ; Åkerud, Tomas LU ; Sterling, A ; Edlund, P-O and Roeraade, J (2003) In Journal of Biomolecular Screening 8(3). p.247-256
Abstract
A method for ligand screening by automated nano-electrospray ionization mass spectrometry (nano-ESI/MS) is described. The core of the system consisted of a chip-based platform for automated sample delivery from a 96-well plate and subsequent analysis based on noncovalent interactions. Human fatty acid binding protein, H-FABP (heart) and A-FABP (adipose), with small potential ligands was analyzed. The technique has been compared with a previously reported method based on nuclear magnetic resonance (NMR), and excellent coorelation with the found hits was obtained. In the current MS screening method, the cycle time per sample was 1.1 min, which is approximately 50 times faster than NMR for single compounds and approximately 5 times faster for... (More)
A method for ligand screening by automated nano-electrospray ionization mass spectrometry (nano-ESI/MS) is described. The core of the system consisted of a chip-based platform for automated sample delivery from a 96-well plate and subsequent analysis based on noncovalent interactions. Human fatty acid binding protein, H-FABP (heart) and A-FABP (adipose), with small potential ligands was analyzed. The technique has been compared with a previously reported method based on nuclear magnetic resonance (NMR), and excellent coorelation with the found hits was obtained. In the current MS screening method, the cycle time per sample was 1.1 min, which is approximately 50 times faster than NMR for single compounds and approximately 5 times faster for compound mixtures. High reproducibility was achieved, and the protein consumption was in the range of 88 to 100 picomoles per sample. Futhermore, a novel protocol for preparation of A-FABP without the natural ligand is presented. The described screening approach is suitable for ligand screening very early in the drug discovery process before conventional high-throughput screens (HTS) are developed and/or used as a secondary screening for ligands identified by HTS. (Less)
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author
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organization
publishing date
type
Contribution to journal
publication status
published
in
Journal of Biomolecular Screening
volume
8
issue
3
pages
247 - 256
publisher
SAGE Publications
external identifiers
  • pmid:12857378
  • wos:000183364000002
  • scopus:0038550485
ISSN
1087-0571
DOI
10.1177/1087057103008003002
language
English
LU publication?
yes
id
7e365256-489b-47c5-bd3c-9a327b1d526e (old id 128015)
date added to LUP
2016-04-01 11:36:42
date last changed
2025-04-03 18:08:17
@article{7e365256-489b-47c5-bd3c-9a327b1d526e,
  abstract     = {{A method for ligand screening by automated nano-electrospray ionization mass spectrometry (nano-ESI/MS) is described. The core of the system consisted of a chip-based platform for automated sample delivery from a 96-well plate and subsequent analysis based on noncovalent interactions. Human fatty acid binding protein, H-FABP (heart) and A-FABP (adipose), with small potential ligands was analyzed. The technique has been compared with a previously reported method based on nuclear magnetic resonance (NMR), and excellent coorelation with the found hits was obtained. In the current MS screening method, the cycle time per sample was 1.1 min, which is approximately 50 times faster than NMR for single compounds and approximately 5 times faster for compound mixtures. High reproducibility was achieved, and the protein consumption was in the range of 88 to 100 picomoles per sample. Futhermore, a novel protocol for preparation of A-FABP without the natural ligand is presented. The described screening approach is suitable for ligand screening very early in the drug discovery process before conventional high-throughput screens (HTS) are developed and/or used as a secondary screening for ligands identified by HTS.}},
  author       = {{Benkestock, K and van Pelt, C K and Åkerud, Tomas and Sterling, A and Edlund, P-O and Roeraade, J}},
  issn         = {{1087-0571}},
  language     = {{eng}},
  number       = {{3}},
  pages        = {{247--256}},
  publisher    = {{SAGE Publications}},
  series       = {{Journal of Biomolecular Screening}},
  title        = {{Automated nano-electrospray mass spectrometry for protein-ligand screening by noncovalent interaction applied to human H-FABP and A-FABP}},
  url          = {{http://dx.doi.org/10.1177/1087057103008003002}},
  doi          = {{10.1177/1087057103008003002}},
  volume       = {{8}},
  year         = {{2003}},
}