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Updated: Jan 4, 2026
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Interactome-Seq: A Protocol for Domainome Library Construction, Validation and Selection by Phage Display and Next Generation Sequencing
Published on: October 3, 2018
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Retooling phage display with electrohydrodynamic nanomixing and nanopore sequencing
Lyndon J Raftery1, Christopher B Howard2, Yadveer S Grewal3
1Australian Institute for Bioengineering and Nanotechnology (AIBN), University of Queensland, Brisbane, Australia. m.trau@uq.edu.au.
Lab on a Chip
|November 13, 2019
Summary
PhageXpress rapidly selects single-chain variable fragment (scFv) molecules for antibody development. This new method significantly accelerates the discovery of bio-recognition molecules like antibodies, reducing screening time and particle usage.
Area of Science:
- Biotechnology
- Immunology
- Molecular Biology
Background:
- Phage display is a powerful technique for isolating single-chain variable fragment (scFv) molecules.
- These scFv molecules can be engineered into therapeutic monoclonal antibodies.
- Traditional phage display methods can be time-consuming and resource-intensive.
Purpose of the Study:
- To develop a rapid and efficient workflow for selecting scFv sequences using phage display.
- To enhance target binding and minimize non-specific interactions during phage library screening.
- To accelerate the discovery of novel bio-recognition molecules for therapeutic and diagnostic applications.
Main Methods:
- PhageXpress workflow utilizing electrohydrodynamic manipulation for enhanced target binding.
- Integration of Oxford Nanopore Technologies' MinION sequencer for high-throughput screening.
- Custom bioinformatics pipeline for analyzing phage library screening data.
- Biopanning against Dengue virus (DENV) non-structural protein 1 (NS1).
Main Results:
- PhageXpress successfully mapped 14 out of 19 anti-NS1 scFv sequences (74%) in a single round of biopanning.
- The new method used approximately 30-fold fewer phage particles compared to traditional methods.
- Traditional methods required 4 weeks and 4 rounds of biopanning to isolate 19 unique scFv clones.
Conclusions:
- PhageXpress significantly accelerates the discovery of scFv molecules compared to traditional phage display methods.
- The developed workflow enhances efficiency by reducing screening time and particle requirements.
- This approach offers a faster route for developing therapeutic and diagnostic biologics.
