Eng Lars, Nygren-Babol Linnéa, Hanning Anders
Episentec, 194 61, Upplands Väsby, Sweden.
Episentec, 194 61, Upplands Väsby, Sweden.
Anal Biochem. 2016 Oct 1;510:79-87. doi: 10.1016/j.ab.2016.06.008. Epub 2016 Jun 17.
Surface plasmon resonance (SPR) is a well-established method for studying interactions between small molecules and biomolecules. In particular, SPR is being increasingly applied within fragment-based drug discovery; however, within this application area, the limited sensitivity of SPR may constitute a problem. This problem can be circumvented by the use of label-enhanced SPR that shows a 100-fold higher sensitivity as compared with conventional SPR. Truly label-free interaction data for small molecules can be obtained by applying label-enhanced SPR in a surface competition assay format. The enhanced sensitivity is accompanied by an increased specificity and inertness toward disturbances (e.g., bulk refractive index disturbances). Label-enhanced SPR can be used for fragment screening in a competitive assay format; the competitive format has the added advantage of confirming the specificity of the molecular interaction. In addition, label-enhanced SPR extends the accessible kinetic regime of SPR to the analysis of very fast fragment binding kinetics. In this article, we demonstrate the working principles and benchmark the performance of label-enhanced SPR in a model system-the interaction between carbonic anhydrase II and a number of small-molecule sulfonamide-based inhibitors.
表面等离子体共振（SPR）是一种成熟的研究小分子与生物分子之间相互作用的方法。特别是，SPR在基于片段的药物发现中应用越来越广泛；然而，在这个应用领域，SPR有限的灵敏度可能会成为一个问题。通过使用标记增强型SPR可以规避这个问题，与传统SPR相比，标记增强型SPR的灵敏度高出100倍。通过在表面竞争测定形式中应用标记增强型SPR，可以获得小分子真正的无标记相互作用数据。增强的灵敏度伴随着特异性的提高以及对干扰（例如，本体折射率干扰）的惰性增加。标记增强型SPR可用于竞争测定形式的片段筛选；竞争形式具有确认分子相互作用特异性的额外优势。此外，标记增强型SPR将SPR可及的动力学范围扩展到非常快速的片段结合动力学分析。在本文中，我们展示了标记增强型SPR的工作原理，并在一个模型系统——碳酸酐酶II与一些基于小分子磺酰胺的抑制剂之间的相互作用中对其性能进行了基准测试。
