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Next-Generation Phage Display to Identify Peptide Ligands of Deubiquitinases

Spiliotopoulos, Anastasios; Maurer, Sigrun K.; Tsoumpeli, Maria T.; Bonfante, Juan A. F.; Owen, Jonathan P.; Gough, Kevin C.; Dreveny, Ingrid


Anastasios Spiliotopoulos

Sigrun K. Maurer

Maria T. Tsoumpeli

Juan A. F. Bonfante

Jonathan P. Owen

Professor of Biochemistry and Pathology


Julie Maupin-Furlow

Mariola J. Edelmann


Phage display (PD) is a powerful method and has been extensively used to generate monoclonal antibodies and identify epitopes, mimotopes, and protein interactions. More recently, the combination of next-generation sequencing (NGS) with PD (NGPD) has revolutionized the capabilities of the method by creating large data sets of sequences from affinity selection-based approaches (biopanning) otherwise challenging to obtain. NGPD can monitor motif enrichment, allow tracking of the selection process over consecutive rounds, and highlight unspecific binders. To tackle the wealth of data obtained, bioinformatics tools have been developed that allow for identifying specific binding sequences (binders) that can then be validated. Here, we provide a detailed account of the use of NGPD experiments to identify ubiquitin-specific protease peptide ligands.


Spiliotopoulos, A., Maurer, S. K., Tsoumpeli, M. T., Bonfante, J. A. F., Owen, J. P., Gough, K. C., & Dreveny, I. (2023). Next-Generation Phage Display to Identify Peptide Ligands of Deubiquitinases. In J. Maupin-Furlow, & M. J. Edelmann (Eds.), Deubiquitinases: Methods and Protocols (189-218). Humana Press.

Online Publication Date Nov 10, 2022
Publication Date 2023
Deposit Date Nov 10, 2022
Publicly Available Date Nov 11, 2024
Pages 189-218
Series Title Methods in Molecular Biology
Series Number 2591
Book Title Deubiquitinases: Methods and Protocols
ISBN 9781071628027
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