Automated Nano-Electrospray Mass Spectrometry for Protein-Ligand Screening by Noncovalent Interaction Applied to Human H-FABP and A-FABP
2003 (English)In: Journal of Biomolecular Screening, ISSN 1087-0571, E-ISSN 1552-454X, Vol. 8, no 3, 247-256 p.Article in journal (Refereed) Published
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 correlation 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. Furthermore, 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.
Place, publisher, year, edition, pages
2003. Vol. 8, no 3, 247-256 p.
automated nano-electrospray; noncovalent; screening; FABP; ACID-BINDING PROTEINS; COMPLEXES; MICRODIALYSIS; RESOLUTION; ADIPOCYTE; RECEPTOR
IdentifiersURN: urn:nbn:se:kth:diva-8358DOI: 10.1177/1087057103008003002ISI: 000183364000002OAI: oai:DiVA.org:kth-8358DiVA: diva2:13659
QC 201007052008-05-072008-05-072010-07-05Bibliographically approved