Astbury Centre for Structural Molecular Biology and the School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, UK.
J Cell Sci. 2022 Jul 15;135(14). doi: 10.1242/jcs.259168. Epub 2022 Jul 18.
Antibodies are the most widely used, traditional tool for labelling molecules in cells. In the past five to ten years, many new labelling tools have been developed with significant advantages over the traditional antibody. Here, we focus on nanobodies and the non-antibody binding scaffold proteins called Affimers. We explain how they are generated, selected and produced, and we describe how their small size, high binding affinity and specificity provides them with many advantages compared to antibodies. Of particular importance, their small size enables them to better penetrate dense cytoskeletal regions within cells, as well as tissues, providing them with specific advantage for super-resolution imaging, as they place the fluorophore with a few nanometres of the target protein being imaged. We expect these novel tools to be of broad interest to many cell biologists and anticipate them becoming the tools of choice for super-resolution imaging.
抗体是最广泛使用的、用于标记细胞内分子的传统工具。在过去五到十年中，已经开发出许多具有明显优于传统抗体的新型标记工具。在这里，我们重点介绍纳米抗体和称为 Affimers 的非抗体结合支架蛋白。我们解释了它们是如何产生、选择和生产的，并描述了它们的小尺寸、高结合亲和力和特异性如何使它们相对于抗体具有许多优势。特别重要的是，它们的小尺寸使它们能够更好地穿透细胞内密集的细胞骨架区域以及组织，为它们提供了用于超分辨率成像的特定优势，因为它们将荧光团放置在离被成像的目标蛋白几个纳米的位置。我们预计这些新型工具将引起许多细胞生物学家的广泛兴趣，并期望它们成为超分辨率成像的首选工具。
