Flemish Institute for Technological Research, VITO, Boeretang 200, B-2400 Mol, Belgium; Institute for Theoretical Physics, KULeuven, Celestijnenlaan 200D, B-3001 Leuven, Belgium.
Institute for Theoretical Physics, KULeuven, Celestijnenlaan 200D, B-3001 Leuven, Belgium.
Biosens Bioelectron. 2015 Feb 15;64:411-5. doi: 10.1016/j.bios.2014.09.043. Epub 2014 Sep 28.
In hybridization based nucleic acid sensors the stringency of hybridization poses a challenge to design and experiment. For a given set of experimental parameters the affinity window of probe-target interaction is always limited and vice versa for a given probe set design, changes in experimental conditions can easily bring some measurements out of detection range. In this paper we introduce and apply a strategy to extend this dynamic range for affinity sensors, sensors which measure the amount of hybridized molecules after equilibrium is reached. The method relies on concepts of additivity of nucleic acids hybridization free energies and on equilibrium isotherms. It consists in combining the measurements from probes with different lengths, by appropriately rescaling the measured signals. We test the validity of the approach on experiments and show that by combining probes with hybridizing regions of length 21, 23 and 25 nucleotides we manage to extend the dynamic range of the intensity signals by a factor of 25. The presented concept is easy to extend, platform free and applies to any hybridization based affinity sensor.
在基于杂交的核酸传感器中，杂交的严谨性对设计和实验提出了挑战。对于给定的一组实验参数，探针-靶标相互作用的亲和窗口总是有限的，反之亦然，对于给定的探针集设计，实验条件的变化很容易使一些测量结果超出检测范围。在本文中，我们引入并应用了一种策略来扩展这种亲和传感器的动态范围，这些传感器测量达到平衡后杂交分子的数量。该方法依赖于核酸杂交自由能的加和性概念和平衡等温线。它包括通过适当缩放测量信号，将来自不同长度探针的测量结果结合起来。我们在实验中验证了该方法的有效性，并表明通过组合杂交区域长度为 21、23 和 25 个核苷酸的探针，我们成功地将强度信号的动态范围扩展了 25 倍。所提出的概念易于扩展，与平台无关，适用于任何基于杂交的亲和传感器。
