Automated Nano-Electrospray Mass Spectrometry for Protein-Ligand Screening by Noncovalent Interaction Applied to Human H-FABP and A-FABP

QC 20100705
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.

NATURVETENSKAP Kemi Analytisk kemi hsv//swe
NATURAL SCIENCES Chemical Sciences Analytical Chemistry hsv//eng
NATURVETENSKAP Kemi Fysikalisk kemi hsv//swe
NATURAL SCIENCES Chemical Sciences Physical Chemistry hsv//eng
automated nano-electrospray; noncovalent; screening; FABP; ACID-BINDING PROTEINS; COMPLEXES; MICRODIALYSIS; RESOLUTION; ADIPOCYTE; RECEPTOR
Analytical chemistry
Analytisk kemi

Van Pelt, C. K.Advion BioSci Inc, Ithaca, NY USA (författare)
Åkerud, TomasLund University,Lunds universitet,Biofysikalisk kemi,Centrum för Molekylär Proteinvetenskap,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Biophysical Chemistry,Center for Molecular Protein Science,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH,Lund Univ, Dept Biophys Chem(Swepub:lu)fkm2-tak (författare)
Sterling, A.Advion BioSci Ltd, Norwich, Norfolk England (författare)
Edlund, Per OlofBiovitrum AB, Analyt Sci (författare)
Roeraade, JohanKTH,Kemi(Swepub:kth)u10i2qgl (författare)
KTHKemi (creator_code:org_t)

Ingår i:Journal of Biomolecular Screening: Elsevier BV8:3, s. 247-2561087-05711552-454X
Ingår i:SLAS Discovery: Elsevier BV8:3, s. 247-2562472-5552

Av författaren/redakt...: Benkestock, Kurt; Van Pelt, C. K.; Åkerud, Tomas; Sterling, A.; Edlund, Per Olof; Roeraade, Johan

Om ämnet

Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.