Thermophoresis in nanoliter droplets to quantify aptamer binding.
Thermophoresis in nanoliter droplets to quantify aptamer binding.
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DOI：
10.1002/anie.201402514
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发表时间：
2014-07
影响因子：
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通讯作者：
S. Seidel;N. Markwardt;Simon A Lanzmich;D. Braun
中科院分区：
文献类型：
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作者：
S. Seidel;N. Markwardt;Simon A Lanzmich;D. Braun
生物分子相互作用是药理学和诊断学的核心。这些相互作用可以通过热电泳（沿着温度梯度的定向分子运动）来量化。它对结合引起的大小、电荷或构象的变化很敏感。已建立的毛细管测量要求每个样品至少0.5 μL。我们将样品消耗减少了50倍，使用声学液滴机器人（Labcyte）生产的10液滴。液滴在油-表面活性剂混合物中稳定，并用红外激光局部加热。荧光显微镜和数值模拟分析了温度升高、马兰戈尼流和浓度分布。在10 nL液滴中，我们量化了amp适体的亲和力、协同性和缓冲剂依赖性。小型化和1536孔板格式使该方法高通量和自动化友好。这促进了人类血清或无标签药物发现筛选诊断分析的创新应用。
Biomolecule interactions are central to pharmacology and diagnostics. These interactions can be quantified by thermophoresis, the directed molecule movement along a temperature gradient. It is sensitive to binding induced changes in size, charge, or conformation. Established capillary measurements require at least 0.5 μL per sample. We cut down sample consumption by a factor of 50, using 10 nL droplets produced with acoustic droplet robotics (Labcyte). Droplets were stabilized in an oil-surfactant mix and locally heated with an IR laser. Temperature increase, Marangoni flow, and concentration distribution were analyzed by fluorescence microscopy and numerical simulation. In 10 nL droplets, we quantified AMP-aptamer affinity, cooperativity, and buffer dependence. Miniaturization and the 1536-well plate format make the method high-throughput and automation friendly. This promotes innovative applications for diagnostic assays in human serum or label-free drug discovery screening.