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Efficient Screening of Combinatorial Peptide Libraries by Spatially Ordered Beads Immobilized on Conventional Glass Slides

1
Division 1.5 Protein Analysis, Federal Institute for Materials Research and Testing (BAM), Richard-Willstätter-Strasse 11, 12489 Berlin, Germany
2
Department of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Straße 2, 12489 Berlin, Germany
3
Institut für Chemie und Biochemie der Freien Universität Berlin, Takustraße 3, 14195 Berlin, Germany
*
Author to whom correspondence should be addressed.
High-Throughput 2019, 8(2), 11; https://doi.org/10.3390/ht8020011
Received: 11 March 2019 / Revised: 8 April 2019 / Accepted: 16 April 2019 / Published: 30 April 2019
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Abstract

Screening of one-bead-one-compound (OBOC) libraries is a proven procedure for the identification of protein-binding ligands. The demand for binders with high affinity and specificity towards various targets has surged in the biomedical and pharmaceutical field in recent years. The traditional peptide screening involves tedious steps such as affinity selection, bead picking, sequencing, and characterization. Herein, we present a high-throughput “all-on-one chip” system to avoid slow and technically complex bead picking steps. On a traditional glass slide provided with an electrically conductive tape, beads of a combinatorial peptide library are aligned and immobilized by application of a precision sieve. Subsequently, the chip is incubated with a fluorophore-labeled target protein. In a fluorescence scan followed by matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF) mass spectrometry, high-affinity binders are directly and unambiguously sequenced with high accuracy without picking of the positive beads. The use of an optimized ladder sequencing approach improved the accuracy of the de-novo sequencing step to nearly 100%. The new technique was validated by employing a FLAG-based model system, identifying new peptide binders for the monoclonal M2 anti-FLAG antibody, and was finally utilized to search for IgG-binding peptides. In the present format, more than 30,000 beads can be screened on one slide. View Full-Text
Keywords: peptide library; screening; HTS; biomarker; on-bead sequencing; on-chip sequencing; microarray; grid; bead array; drug screening; lead compounds; pharmaceutical development; affinity reagents peptide library; screening; HTS; biomarker; on-bead sequencing; on-chip sequencing; microarray; grid; bead array; drug screening; lead compounds; pharmaceutical development; affinity reagents
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Schwaar, T.; Lettow, M.; Remmler, D.; Börner, H.G.; Weller, M.G. Efficient Screening of Combinatorial Peptide Libraries by Spatially Ordered Beads Immobilized on Conventional Glass Slides. High-Throughput 2019, 8, 11.

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