Comparison of High-Throughput Sequencing for Phage Display Peptide Screening on Two Commercially Available Platforms

Abstract

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.