Department of Inorganic and Analytical Chemistry, University of Debrecen, Egyetem ter 1., Debrecen, 4032, Hungary.
Department of Inorganic and Analytical Chemistry, University of Debrecen, Egyetem ter 1., Debrecen, 4032, Hungary.
Anal Chim Acta. 2020 Apr 29;1108:70-78. doi: 10.1016/j.aca.2020.02.048. Epub 2020 Feb 26.
This paper demonstrates the design, efficiency and applicability of a simple and inexpensive microfluidic immobilized enzymatic reactor (IMER) for rapid protein digestion. The high surface-to-volume ratio (S/V) of the reactor was achieved by forming pillars in the channel. It was found that pillar arrays including dimensions of 40 μm × 40 μm as pillar diameter and interpillar distance can provide both relatively high S/V and flow rate in the PDMS chip, the fabrication of which was performed by means of soft lithography using average research laboratory infrastructure. CZE peptide maps of IMER-based digestions were compared to peptide maps obtained from standard in-solution digestion of proteins. The peak patterns of the electropherograms and the identified proteins were similar, however, digestion with the IMER requires less than 10 min, while in-solution digestion takes 16 h.
本文展示了一种简单且廉价的微流控固定化酶反应器 (IMER) 的设计、效率和适用性，该反应器可用于快速蛋白质消化。通过在通道中形成柱子，实现了高的比表面积（S/V）。研究发现，包括直径为 40 μm×40 μm 的柱子直径和柱子之间的距离的柱子阵列，可以在 PDMS 芯片中提供相对较高的 S/V 和流速，该 PDMS 芯片是通过使用平均研究实验室基础设施的软光刻技术制造的。基于 IMER 的消化的 CZE 肽图谱与从蛋白质的标准溶液消化获得的肽图谱进行了比较。电泳图谱的峰型和鉴定的蛋白质相似，然而，使用 IMER 进行消化需要不到 10 分钟，而溶液消化需要 16 小时。
