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Article Dans Une Revue Scientific Reports Année : 2016

A rapid and practical technique for real-time monitoring of biomolecular interactions using mechanical responses of macromolecules

Résumé

Monitoring biological reactions using the mechanical response of macromolecules is an alternative approach to immunoassays for providing real-time information about the underlying molecular mechanisms. Although force spectroscopy techniques, e.g. AFM and optical tweezers, perform precise molecular measurements at the single molecule level, sophisticated operation prevent their intensive use for systematic biosensing. Exploiting the biomechanical assay concept, we used micro-electro mechanical systems (MEMS) to develop a rapid platform for monitoring bio/chemical interactions of bio macromolecules, e.g. DNA, using their mechanical properties. The MEMS device provided real-time monitoring of reaction dynamics without any surface or molecular modifications. A microfluidic device with a side opening was fabricated for the optimal performance of the MEMS device to operate at the air-liquid interface for performing bioassays in liquid while actuating/sensing in air. The minimal immersion of the MEMS device in the channel provided long-term measurement stability (>10 h). Importantly, the method allowed monitoring effects of multiple solutions on the same macromolecule bundle (demonstrated with DNA bundles) without compromising the reproducibility. We monitored two different types of effects on the mechanical responses of DNA bundles (stiffness and viscous losses) exposed to pH changes (2.1 to 4.8) and different Ag + concentrations (1 μM to 0.1 M). Mechanical properties of molecules are used to investigate a wide range of biological and chemical mechanisms varying from nucleic acid conformation to enzymatic reaction kinetics and from cell manipulation to motor protein operation 1. Unlike some of the other well-established methods that are often limited to smaller sized molecules , e.g. nuclear magnetic resonance (NMR) 2 and surface plasmon resonance (SPR) 3 , force spectroscopy techniques , e.g. optical tweezers 4,5 , magnetic tweezers 6-8 and AFM 9,10 , provide sensitive measurements on mechanical properties spanning six orders of magnitude in length (10 −10-10 −4 m) including macromolecules 1,11,12. These techniques provide information at the single molecule level to reveal novel properties of various different molecules. However, low throughput, surface modification, high-level operational skills and complicated calibra-tion/setup procedures exclude these techniques for rapid routine tests that are essential to provide statistically significant biomechanical information especially for clinical studies 13. As a result, a rapid, practical, time-and cost-efficient, automated and preferably portable method is beneficial as a complementary or alternative method to the conventional techniques to be employed even by non-specialists. MEMS technology presents certain advantages to cover the mentioned requirements for routinely performed assays on mechanical responses of macromolecules. MEMS, capable of conducting real-time mechanical sensing with electrical readouts associated with low noise and high stability measurements 14 , allows low
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Dates et versions

hal-02105866 , version 1 (22-04-2019)

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Mehmet Tarhan, Nicolas Lafitte, Yannick Tauran, Laurent Jalabert, Momoko Kumemura, et al.. A rapid and practical technique for real-time monitoring of biomolecular interactions using mechanical responses of macromolecules. Scientific Reports, 2016, 6 (1), ⟨10.1038/srep28001⟩. ⟨hal-02105866⟩
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