{“_”：“通过单分子置换统计的溶液相生物分子反应和相互作用动力学。”，“i”：[“原位”]}
{"_":" Kinetics of Solution-Phase Biomolecular Reactions and Interactions through Single-Molecule Displacement Statistics.","i":["In Situ"]}
摘要(中文译文)
我们报告了一个通过单分子位移统计来量化溶液相中生物（大）分子反应动力学的框架。利用单分子位移/扩散率映射（SMM），我们在宽视场中以超高吞吐量测量自由扩散分子的瞬态（0.6毫秒）位移。将许多分子的时间依赖位移分布拟合到双组分扩散模型，基于分子量差异量化反应物和产物比例，从而能够以约1秒的时间分辨率对反应进程进行动力学分析。在约100 pM荧光标记反应物条件下，通过在大范围内改变第二种未标记反应物的浓度，我们测定了应变促进叠氮-炔环加成和N-羟基琥珀酰亚胺酯氨解的二阶反应速率常数，以及后者的一阶水解副反应速率常数。对于抗体结合动力学，我们量化了单克隆抗体的结合和解离速率常数，并揭示了多克隆抗体交联复合物/聚集体的逐渐形成。在微升规模样品条件下，我们基于单分子位移的方法提供了与分子大小相关的直观读数，并适用于多种反应类型和条件。
Abstract
We report a framework for quantifying the reaction kinetics of bio-(macro)-molecules in the solution phase through single-molecule displacement statistics. Using single-molecule displacement/diffusivity mapping (SMM), we measure the transient (0.6 ms) displacements of freely diffusing molecules in the wide field at ultrahigh throughput. Fitting the time-dependent displacement distributions of many molecules to a two-component diffusion model quantifies the reactant and product fractions based on molecular-weight differences, thereby enabling kinetic analysis of reaction progress at ∼1 s temporal resolution. Working with ∼100 pM fluorescently tagged reactant while varying the concentration of a second, unlabeled reactant over wide ranges, we determine second-order reaction rate constants for strain-promoted azide-alkyne cycloaddition and -hydroxysuccinimide ester aminolysis as well as first-order hydrolysis side-reaction rate constants for the latter. For antibody binding kinetics, we quantify association and dissociation rate constants for a monoclonal antibody and unveil the gradual formation of cross-linked complexes/aggregates for a polyclonal antibody. Working with microliter-scale samples, our single-molecule-displacement-based approach provides intuitive readouts linked to the molecular size and is applicable to diverse reaction types and conditions.
如何引用
AMA
Wan Li, Ke Xu. {"_":" Kinetics of Solution-Phase Biomolecular Reactions and Interactions through Single-Molecule Displacement Statistics.","i":["In Situ"]}. ACS central science. 2026; doi:10.1021/acscentsci.6c00063.
APA
Wan Li, Ke Xu (2026). {"_":" Kinetics of Solution-Phase Biomolecular Reactions and Interactions through Single-Molecule Displacement Statistics.","i":["In Situ"]}. ACS central science. https://doi.org/10.1021/acscentsci.6c00063
GB/T 7714
Wan Li, Ke Xu. {"_":" Kinetics of Solution-Phase Biomolecular Reactions and Interactions through Single-Molecule Displacement Statistics.","i":["In Situ"]}[J]. ACS central science, 2026 doi:10.1021/acscentsci.6c00063.
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