V R Eugene Christo, Kloth Esther Charlotte Sophia, Nisini Filippo, Reuther Cordula, Diez Stefan
B CUBE - Center for Molecular Bioengineering, TUD Dresden University of Technology, 01307 Dresden, Germany.
Cluster of Excellence Physics of Life, TUD Dresden University of Technology, 01062, Dresden, Germany.
Nano Lett. 2025 May 21;25(20):8194-8202. doi: 10.1021/acs.nanolett.5c01188. Epub 2025 May 8.
Microtubule gliding assays provide a unique mechanism for molecular detection in which binding of analytes to the microtubule lattice reduces the microtubule gliding speed. The reduction in the gliding speed correlates with the density of the bound analytes, enabling its quantification. Although promising, this technique is still in the proof-of-concept stage. Improving the sensitivity and limit of detection of the assay could make the technique comparable to that of advanced molecular detection methods. This study demonstrates that reducing the ionic strength of the buffer increases the sensitivity of the assay by enhancing the interactions between kinesin and microtubules. When using a low ionic strength buffer (BRB10) compared with a standard buffer (BRB80), we observed a more pronounced reduction in microtubule gliding speed in the presence of analytes, improving the detection limit. Therefore, this approach offers a simple and scalable way to improve the sensitivity of motor-based detection assays.
微管滑动分析提供了一种独特的分子检测机制，其中分析物与微管晶格的结合会降低微管的滑动速度。滑动速度的降低与结合的分析物密度相关，从而能够对其进行定量。尽管很有前景，但这项技术仍处于概念验证阶段。提高该分析方法的灵敏度和检测限可使该技术与先进的分子检测方法相媲美。本研究表明，降低缓冲液的离子强度可通过增强驱动蛋白与微管之间的相互作用来提高分析的灵敏度。与标准缓冲液（BRB80）相比，当使用低离子强度缓冲液（BRB10）时，我们观察到在存在分析物的情况下微管滑动速度有更明显的降低，从而提高了检测限。因此，这种方法提供了一种简单且可扩展的方式来提高基于运动蛋白的检测分析的灵敏度。
