Comparison of High-Throughput Sequencing for Phage Display Peptide Screening on Two Commercially Available Platforms
Phage display is a widely used technique to screen peptide sequences for interaction with target biomolecules, such as proteins and cells. Traditional protocols screen for only a limited fraction of a candidate pool due to the cost and time limitation of Sanger sequencing. Recent developments in high-throughput sequencing (HTS) technologies enable researchers to assess millions of biomolecule sequences. In this study, eluted DNA pooled from a phage display screening were sequenced by two types of HTS methodologies; sequence by synthesis and a nanopore-based techniques. While both methods resulted in the identification of several candidate peptide motifs determined to be interacting with the target proteins, the sequence by synthesis method provide a higher yield of valid reads for data analyses. Input library preparation protocol for the nanopore sequencer needs further modification to achieve effective data collection.
KeywordsPeptide motifs Phage display Next-generation sequencing Bioinformatics Protein–protein interaction
I would like to thank Michigan Technological University Research Excellence Fund (R01547) and Portage Health Foundation for funding (R75389).
Compliance with ethical standards
Conflict of interest
The author declares no conflict of interest.
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