Rob Tamanna, Wilson Derek J
Center for Research in Mass Spectrometry, Department of Chemistry, York University, Toronto, Ontario, Canada.
J Am Soc Mass Spectrom. 2009 Jan;20(1):124-30. doi: 10.1016/j.jasms.2008.09.005. Epub 2008 Sep 7.
An electrospray coupled microfluidic reactor for the measurement of millisecond time-scale, solution phase kinetics is introduced. The device incorporates a simple two-channel design that is etched into polymethyl methacrylate (PMMA) by laser ablation. The outlet of the device is laser cut to a sharp tip, facilitating low dead volume 'on chip' electrospray. Fabrication is fast, straightforward and highly reproducible, supporting rapid prototyping and large-scale reproduction. Device performance is characterized using a cytochrome c unfolding reaction. Unfolding processes with rates in excess of 30 s(-1) are easily measured, including the appearance of a 'native-like' intermediate that is maximally populated 180 ms post reaction initiation. To extract reliable rates from the data, a theoretical framework for the analysis of kinetics acquired under square-channel laminar flow is introduced.
介绍了一种用于测量毫秒时间尺度溶液相动力学的电喷雾耦合微流控反应器。该装置采用简单的双通道设计，通过激光烧蚀蚀刻到聚甲基丙烯酸甲酯（PMMA）中。该装置的出口经激光切割成尖锐的尖端，便于进行低死体积的“芯片上”电喷雾。制造过程快速、直接且高度可重复，支持快速原型制作和大规模复制。使用细胞色素c的去折叠反应来表征装置性能。可以轻松测量速率超过30 s⁻¹的去折叠过程，包括出现一个“类天然”中间体，该中间体在反应开始后180毫秒时达到最大丰度。为了从数据中提取可靠的速率，引入了一个用于分析在方通道层流条件下获得的动力学的理论框架。
