Pitfalls to avoid when using phage display for snake toxins

Andreas Hougaard Laustsen; Line Præst Lauridsen; Bruno Lomonte; Mikael Rørdam Andersen; Brian Lohse

doi:10.1016/j.toxicon.2016.12.010

Pitfalls to avoid when using phage display for snake toxins

Andreas Hougaard Laustsen, Line Præst Lauridsen, Bruno Lomonte, Mikael Rørdam Andersen, Brian Lohse

6 Citations (Scopus)

Abstract

Antivenoms against bites and stings from snakes, spiders, and scorpions are associated with immunological side effects and high cost of production, since these therapies are still derived from the serum of hyper-immunized production animals. Biotechnological innovations within envenoming therapies are thus warranted, and phage display technology may be a promising avenue for bringing antivenoms into the modern era of biologics. Although phage display technology represents a robust and high-throughput approach for the discovery of antibody-based antitoxins, several pitfalls may present themselves when animal toxins are used as targets for phage display selection. Here, we report selected critical challenges from our own phage display experiments associated with biotinylation of antigens, clone picking, and the presence of amber codons within antibody fragment structures in some phage display libraries. These challenges may be detrimental to the outcome of phage display experiments, and we aim to help other researchers avoiding these pitfalls by presenting their solutions.

Original language	English
Journal	Toxicon
Volume	126
Pages (from-to)	79-89
Number of pages	11
ISSN	0041-0101
DOIs	https://doi.org/10.1016/j.toxicon.2016.12.010
Publication status	Published - 1 Feb 2017

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title = "Pitfalls to avoid when using phage display for snake toxins",

abstract = "Antivenoms against bites and stings from snakes, spiders, and scorpions are associated with immunological side effects and high cost of production, since these therapies are still derived from the serum of hyper-immunized production animals. Biotechnological innovations within envenoming therapies are thus warranted, and phage display technology may be a promising avenue for bringing antivenoms into the modern era of biologics. Although phage display technology represents a robust and high-throughput approach for the discovery of antibody-based antitoxins, several pitfalls may present themselves when animal toxins are used as targets for phage display selection. Here, we report selected critical challenges from our own phage display experiments associated with biotinylation of antigens, clone picking, and the presence of amber codons within antibody fragment structures in some phage display libraries. These challenges may be detrimental to the outcome of phage display experiments, and we aim to help other researchers avoiding these pitfalls by presenting their solutions.",

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T1 - Pitfalls to avoid when using phage display for snake toxins

AU - Laustsen, Andreas Hougaard

AU - Lauridsen, Line Præst

AU - Lomonte, Bruno

AU - Andersen, Mikael Rørdam

AU - Lohse, Brian

PY - 2017/2/1

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N2 - Antivenoms against bites and stings from snakes, spiders, and scorpions are associated with immunological side effects and high cost of production, since these therapies are still derived from the serum of hyper-immunized production animals. Biotechnological innovations within envenoming therapies are thus warranted, and phage display technology may be a promising avenue for bringing antivenoms into the modern era of biologics. Although phage display technology represents a robust and high-throughput approach for the discovery of antibody-based antitoxins, several pitfalls may present themselves when animal toxins are used as targets for phage display selection. Here, we report selected critical challenges from our own phage display experiments associated with biotinylation of antigens, clone picking, and the presence of amber codons within antibody fragment structures in some phage display libraries. These challenges may be detrimental to the outcome of phage display experiments, and we aim to help other researchers avoiding these pitfalls by presenting their solutions.

AB - Antivenoms against bites and stings from snakes, spiders, and scorpions are associated with immunological side effects and high cost of production, since these therapies are still derived from the serum of hyper-immunized production animals. Biotechnological innovations within envenoming therapies are thus warranted, and phage display technology may be a promising avenue for bringing antivenoms into the modern era of biologics. Although phage display technology represents a robust and high-throughput approach for the discovery of antibody-based antitoxins, several pitfalls may present themselves when animal toxins are used as targets for phage display selection. Here, we report selected critical challenges from our own phage display experiments associated with biotinylation of antigens, clone picking, and the presence of amber codons within antibody fragment structures in some phage display libraries. These challenges may be detrimental to the outcome of phage display experiments, and we aim to help other researchers avoiding these pitfalls by presenting their solutions.

U2 - 10.1016/j.toxicon.2016.12.010

DO - 10.1016/j.toxicon.2016.12.010

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Pitfalls to avoid when using phage display for snake toxins

Abstract

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