Retooling phage display with electrohydrodynamic nanomixing and nanopore sequencing
Lyndon J Raftery, Christopher B Howard, Yadveer S Grewal, Ramanathan Vaidyanathan, Martina L Jones, Will Anderson, Darren Korbie, Tania Duarte, Duc Cao Minh, Hoang Nguyen Son, Lachlan JM Coin, Stephen M Mahler, Matt Trau
LAB ON A CHIP | ROYAL SOC CHEMISTRY | Published : 2019
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 (NS..View full abstract
Awarded by National Breast Cancer Foundation of Australia
Although not directly funding the research work in this paper, we acknowledge the funding received by our laboratory from the National Breast Cancer Foundation of Australia (CG-12-07). This grant has significantly contributed to the environment to stimulate the research described here.