Open AccessDevelopment of a New Approach for Screening Combinatorial Libraries Using MALDI-TOF-MS and HPLC-ESI-MS/MS
Author(s) -
Sonal Mathur,
Marc Hassel,
Frank Steiner,
Klaus Hollemeyer,
Rolf W. Hartmann
Publication year - 2003
Publication title -
slas discovery
Language(s) - English
Resource type - Journals
eISSN - 2472-5560
pISSN - 2472-5552
DOI - 10.1177/1087057103253333
Subject(s) - chemistry , chromatography , peptide library , peptide , high performance liquid chromatography , mass spectrometry , high throughput screening , aldehyde , combinatorial chemistry , peptide sequence , biochemistry , gene , catalysis
Mass spectrometric techniques play a prominent role in the rapidly expanding field of high-throughput screening (HTS). In this paper, the authors present a novel qualitative approach for the screening of a small library of compounds using MALDI-TOF-MS and HPLC-ESI-MS/MS. Chymotrypsin (CT), a serine protease, was selected as the target protein. A well-known inhibitor of CT is chymostatin (CS), a naturally occurring peptide aldehyde, which is reported to be a mixture of 3 components—A, B, and C—differing only in one of the amino acids. The authors report that native CS mixture consists of 3 additional ring hydroxylated components and that each compound exists in 2 epimeric forms. In case of protein-binding compounds, only 1 of the epimers was found to be active. A unique feature of this study is the generation of a combinatorial library of CS derivatives applying a one-pot strategy followed by identification and structural elucidation of the library components. Analytical investigation of the library resulted in the identification of 22 compounds. After incubation with CT and centrifugal ultrafiltration, 10 compounds were detected as protein-binding ligands. Finally, the complementary potentials of MALDI-TOF-MS and HPLC-MS/MS in the screening of complex ligand mixtures have been discussed. ( Journal of Biomolecular Screening 2003:136-148)