Retooling phage display with electrohydrodynamic nanomixing and nanopore sequencing

Phage display methodologies offer a versatile platform for the isolation of single-chain Fv (scFv) molecules which may be rebuilt into monoclonal antibodies. Herein, we report on a complete workflow termed PhageXpress, for rapid selection of single-chain Fv sequences by leveraging electrohydrodynami...

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Bibliographic Details
Published in:Lab on a chip 2019-12, Vol.19 (24), p.483-492
Main Authors: Raftery, Lyndon J, Howard, Christopher B, Grewal, Yadveer S, Vaidyanathan, Ramanathan, Jones, Martina L, Anderson, Will, Korbie, Darren, Duarte, Tania, Cao, Minh Duc, Nguyen, Son Hoang, Coin, Lachlan J. M, Mahler, Stephen M, Trau, Matt
Format: Article
Language:English
Subjects:
Antigens
Bioinformatics
Chains
Diagnostic systems
Electrohydrodynamics
Monoclonal antibodies
Phages
Porosity
Screening
Viral diseases
Viruses
Workflow
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Summary:Phage display methodologies offer a versatile platform for the isolation of single-chain Fv (scFv) molecules which may be rebuilt into monoclonal antibodies. Herein, we report on a complete workflow termed PhageXpress, for rapid selection of single-chain Fv sequences by leveraging electrohydrodynamic-manipulation of a solution containing phage library particles to enhance target binding whilst minimizing non-specific interactions. Our PhageXpress technique is combined with Oxford Nanopore Technologies' MinION sequencer and custom bioinformatics to achieve high-throughput screening of phage libraries. We performed 4 rounds of biopanning against Dengue virus (DENV) non-structural protein 1 (NS1) using traditional methods (4 week turnaround), which resulted in the isolation of 19 unique scFv clones. We validated the feasibility and efficiency of the PhageXpress method utilizing the same phage library and antigen target. Notably, we successfully mapped 14 of the 19 anti-NS1 scFv sequences (∼74%) with our new method, despite using ∼30-fold less particles during screening and conducting only a single round of biopanning. We believe this approach supersedes traditional methods for the discovery of bio-recognition molecules such as antibodies by speeding up the process for the development of therapeutic and diagnostic biologics. High throughput screening of phage display libraries for target binding molecules using electrohydrodynamic nanomixing and nanopore sequencing.
ISSN:1473-0197
1473-0189
DOI:10.1039/c9lc00978g