Department of Engineering, Gustav Wieds Vej 10, Aarhus University, 8000 Aarhus C, Denmark.
Sci Rep. 2017 Feb 10;7:42230. doi: 10.1038/srep42230.
One of the hallmarks of cancer is sustained angiogenesis. Here, normal endothelial cells are activated, and their formation of new blood vessels leads to continued tumour growth. An improved patient condition is often observed when angiogenesis is prevented or normalized through targeting of these genomically stable endothelial cells. However, intracellular targets constitute a challenge in therapy, as the agents modulating these targets have to be delivered and internalized specifically to the endothelial cells. Selection of antibodies binding specifically to certain cell types is well established. It is nonetheless a challenge to ensure that the binding of antibodies to the target cell will mediate internalization. Previously selection of such antibodies has been performed targeting cancer cell lines; most often using either monovalent display or polyvalent display. In this article, we describe selections that isolate internalizing antibodies by sequential combining monovalent and polyvalent display using two types of helper phages, one which increases display valence and one which reduces background. One of the selected antibodies was found to mediate internalization into human endothelial cells, although our results confirms that the single stranded nature of the DNA packaged into phage particles may limit applications aimed at targeting nucleic acids in mammalian cells.
癌症的一个特征是持续的血管生成。在这里，正常的内皮细胞被激活，它们形成新的血管，导致肿瘤持续生长。通过针对这些基因组稳定的内皮细胞来阻止或使血管正常化，可以观察到患者病情的改善。然而，细胞内靶点在治疗中构成了挑战，因为调节这些靶点的药物必须被递送到内皮细胞并被特异性内化。结合特定于某些细胞类型的抗体的选择已经得到很好的确立。然而，确保抗体与靶细胞的结合将介导内化仍然是一个挑战。以前，针对癌细胞系进行了这种抗体的选择；最常用的方法是单价展示或多价展示。在本文中，我们描述了通过使用两种辅助噬菌体进行顺序组合单价和多价展示来分离内化抗体的选择，一种增加展示效价，一种降低背景。其中一种选择的抗体被发现可以将内吞作用介导到人类内皮细胞中，尽管我们的结果证实，包装在噬菌体颗粒中的单链 DNA 的性质可能限制了针对哺乳动物细胞中核酸的靶向应用。
