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MALDI-Supported Screening of Large Peptide Libraries for Identification of Protein Binders

  • Screening of one-bead-one-peptide libraries is a powerful analytical tool for the identification of protein ligands. However, the traditional peptide screening procedure involves tedious steps such as manual selection, sequencing, and characterization. We present a high-throughput “all in one chip” system, allowing the screening of a high number of resin beads in short time. Here, beads of a combinatorial one-bead one compound peptide library are immobilized on an in-house produced chip, on which every bead has a well-defined position. The chip is then incubated with a fluorophore-labeled protein, identifying suitable peptides by a high-resolution fluorescence scan. The screening is followed by MALDI-MS experiments directly on the respective glass chip. To circumvent the need for peptide fragmentation normally used for peptide de novo sequencing, which can result in incomplete sequence information, an approach based on ladder sequencing has been used. This allows the peptide sequenceScreening of one-bead-one-peptide libraries is a powerful analytical tool for the identification of protein ligands. However, the traditional peptide screening procedure involves tedious steps such as manual selection, sequencing, and characterization. We present a high-throughput “all in one chip” system, allowing the screening of a high number of resin beads in short time. Here, beads of a combinatorial one-bead one compound peptide library are immobilized on an in-house produced chip, on which every bead has a well-defined position. The chip is then incubated with a fluorophore-labeled protein, identifying suitable peptides by a high-resolution fluorescence scan. The screening is followed by MALDI-MS experiments directly on the respective glass chip. To circumvent the need for peptide fragmentation normally used for peptide de novo sequencing, which can result in incomplete sequence information, an approach based on ladder sequencing has been used. This allows the peptide sequence identification by fragmentation-free MS with almost 100 % accuracy. For this purpose, a software tool was developed automatically translating MALDI-MS spectra into the corresponding peptide sequences.zeige mehrzeige weniger

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Metadaten
Autoren/innen:Timm Schwaar
Koautoren/innen:Michael G. Weller, Maike Lettow, Dario Remmler, Hans G. Börner
Dokumenttyp:Posterpräsentation
Veröffentlichungsform:Präsentation
Sprache:Englisch
Jahr der Erstveröffentlichung:2019
Organisationseinheit der BAM:1 Analytische Chemie; Referenzmaterialien
1 Analytische Chemie; Referenzmaterialien / 1.5 Proteinanalytik
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Anorganische Chemie
Freie Schlagwörter:Combinatorial screening; OBOC; Peptide library
Themenfelder/Aktivitätsfelder der BAM:Analytical Sciences
Analytical Sciences / Spurenanalytik und chemische Zusammensetzung
Veranstaltung:ANAKON
Veranstaltungsort:Münster, Germany
Beginndatum der Veranstaltung:25.03.2019
Enddatum der Veranstaltung:28.03.2019
Verfügbarkeit des Dokuments:Datei im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:10.04.2019
Referierte Publikation:Nein