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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Published in | Lab on a chip Vol. 19; no. 24; pp. 483 - 492 |
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Main Authors | , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
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England
Royal Society of Chemistry
21.12.2019
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Abstract | 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. |
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AbstractList | 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. 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. |
Author | Trau, Matt Korbie, Darren Coin, Lachlan J. M Mahler, Stephen M Howard, Christopher B Vaidyanathan, Ramanathan Anderson, Will Raftery, Lyndon J Cao, Minh Duc Jones, Martina L Grewal, Yadveer S Duarte, Tania Nguyen, Son Hoang |
AuthorAffiliation | Centre for Personalised Nanomedicine ARC Training Centre for Biopharmaceutical Innovation Australian Institute for Bioengineering and Nanotechnology (AIBN) University of Queensland AIBN Institute for Molecular Bioscience School of Chemistry and Molecular Biosciences (SCMB) |
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Author_xml | – sequence: 1 givenname: Lyndon J surname: Raftery fullname: Raftery, Lyndon J – sequence: 2 givenname: Christopher B surname: Howard fullname: Howard, Christopher B – sequence: 3 givenname: Yadveer S surname: Grewal fullname: Grewal, Yadveer S – sequence: 4 givenname: Ramanathan surname: Vaidyanathan fullname: Vaidyanathan, Ramanathan – sequence: 5 givenname: Martina L surname: Jones fullname: Jones, Martina L – sequence: 6 givenname: Will surname: Anderson fullname: Anderson, Will – sequence: 7 givenname: Darren surname: Korbie fullname: Korbie, Darren – sequence: 8 givenname: Tania surname: Duarte fullname: Duarte, Tania – sequence: 9 givenname: Minh Duc surname: Cao fullname: Cao, Minh Duc – sequence: 10 givenname: Son Hoang surname: Nguyen fullname: Nguyen, Son Hoang – sequence: 11 givenname: Lachlan J. M surname: Coin fullname: Coin, Lachlan J. M – sequence: 12 givenname: Stephen M surname: Mahler fullname: Mahler, Stephen M – sequence: 13 givenname: Matt surname: Trau fullname: Trau, Matt |
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SubjectTerms | Antigens Bioinformatics Chains Diagnostic systems Electrohydrodynamics Monoclonal antibodies Phages Porosity Screening Viral diseases Viruses Workflow |
Title | Retooling phage display with electrohydrodynamic nanomixing and nanopore sequencing |
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