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Updated: May 6, 2026

A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits

A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits

Published on: February 16, 2014

Affinity selection using filamentous phage display.

Mari Ann Kulseth¹, Annette Fagerlund, Astrid Hilde Myrset

¹Department of Medical Genetics, Rikshospitalet, Oslo University Hospital, Oslo, Norway.

Methods in Molecular Biology (Clifton, N.J.)

|October 23, 2013

Summary

Phage display is a powerful method for discovering peptide binders for therapeutic targets. This study ensures selection accuracy by controlling phage propagation during affinity selection processes.

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Area of Science:

Biotechnology
Molecular Biology
Immunology

Background:

Phage display is an in vitro selection technique used to identify peptide binders for protein targets.
Successful identification of therapeutic peptide binders relies on accurate affinity selection, free from biases in phage propagation rates.

Purpose of the Study:

To present methods for protein target presentation in phage display.
To provide a protocol for phage rescue and propagation with controls to ensure unbiased selection.

Main Methods:

Utilizing filamentous phage for peptide display.
Implementing affinity selection of phage libraries against specific protein targets.
Employing controlled phage rescue and propagation protocols with built-in checks.

Main Results:

Demonstrated two distinct approaches for presenting protein targets during phage display selection.
Established a robust protocol for phage rescue and propagation that mitigates selection bias.
Introduced controls to ensure equal conditions for eluted phages during amplification.

Conclusions:

The presented methods and protocols enhance the reliability of phage display for identifying high-affinity peptide binders.
Proper surveillance of the phage life cycle is critical for successful affinity selection and therapeutic target identification.