Laustsen Andreas Hougaard, Lauridsen Line Præst, Lomonte Bruno, Andersen Mikael Rørdam, Lohse Brian
Department of Biotechnology and Biomedicine, Technical University of Denmark, Denmark; Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.
Department of Biotechnology and Biomedicine, Technical University of Denmark, Denmark.
Toxicon. 2017 Feb;126:79-89. doi: 10.1016/j.toxicon.2016.12.010. Epub 2016 Dec 23.
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.
针对蛇、蜘蛛和蝎子咬伤及蜇伤的抗蛇毒血清存在免疫副作用且生产成本高昂，因为这些疗法仍来源于超免疫生产动物的血清。因此，毒液疗法的生物技术创新很有必要，噬菌体展示技术可能是将抗蛇毒血清带入现代生物制剂时代的一条有前景的途径。尽管噬菌体展示技术是发现基于抗体的抗毒素的一种强大且高通量的方法，但当动物毒素用作噬菌体展示筛选的靶标时，可能会出现一些问题。在此，我们报告了我们自己的噬菌体展示实验中与抗原生物素化、克隆挑选以及某些噬菌体展示文库中抗体片段结构内琥珀密码子的存在相关的一些关键挑战。这些挑战可能对噬菌体展示实验的结果不利，我们旨在通过给出解决方案来帮助其他研究人员避免这些陷阱。
