Pitfalls to avoid when using phage display for snake toxins

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• 作者：Andreas Hougaard Laustsen^a ; ^b ; Line Præ ; st Lauridsen^a ; Bruno Lomonte^c ; Mikael Rø ; rdam Andersen^a ; Brian Lohse^b ; ^{bril@sund.ku.dk}
• 关键词：Recombinant antivenom ; Next generation antivenom ; Phage display ; Biotinylation ; Antibody technology ; Amber codons ; Clone picking
• 刊名：Toxicon
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：126
• 期：Complete
• 页码：79-89
• 全文大小：1837 K
• 卷排序：126

文摘

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.