Interaction Analysis through Proteomic Phage Display

Figure 1

Schematic representation of proteomic phage display using the M13 phage. Input phage display libraries are constructed from cDNA, ORFs, or oligonucleotide arrays designed from a proteome of interest (1). Peptides are displayed on pVIII (2). Bait proteins are immobilized on a solid surface and incubated with the naïve input phage library (3). Binding of phage occurs through interactions between displayed peptides and bait proteins, but nonspecific interactions cause noise in the selection (not shown). Unbound phage is washed away (4) and bound phage is eluted through acidic or basic conditions or by the addition of actively growing host bacteria (5). Eluted phage is amplified (6) and used for repeated (typically 3–5) cycles of selection, which is necessary to amplify specifically bound phage over nonspecific binders. Sanger sequencing of confirmed binders and/or NGS of the retained phage pools provides lists of binders from the target proteome (7).