微流控圆盘上的微型化和高度平行的蛋白质结晶

C. Steinert, J. Mueller-Dieckmann, M. Weiss, M. Roessle, R. Zengerle, P. Koltay
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引用次数: 9

摘要

本文首次采用自由界面扩散(FID)方法,提出了一种新型的微流体系统,用于微型化和高度平行的蛋白质结晶实验。该新型系统基于热压成型的微流控圆盘,该圆盘具有100个蛋白质结晶室,可并行进行多达100种不同的结晶实验。微观结构的最小特征尺寸为30mm,最大纵横比为1。圆盘的流体设计可以在最小体积为5nl的结晶室中分层纳升体积的蛋白质和结晶溶液(沉淀剂)。蛋白质样品通过非接触式nL分配器加载到光盘上,最小剂量体积为1nl,死亡体积仅为500nl。圆盘上的所有液体处理步骤都是由圆盘旋转引起的离心力完成的。高达80mum的大晶体过氧化氢酶,溶菌酶,蛋白酶K和胰岛素,已在光盘上生产,以证明适当的性能。随后,在x射线实验中对晶体进行原位分析,而不将其从光盘中取出。
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Miniaturized and highly parallel protein crystallization on a microfluidic disc
For the first time we present a new microfluidic system for miniaturized and highly parallel protein crystallization experiments by the free interface diffusion (FID) method. The novel system is based on a microfluidic disc fabricated by hot embossing which features 100 protein crystallization chambers enabling up to 100 different crystallization experiments in parallel. The mi- crostructures exhibit minimal feature sizes of 30mum and a maximum aspect ratio of 1. The fluidic design of the disc enables lamination of nanoliter volumes of protein and crystallization solution (precipitant) in a crystallization chamber of minimum volume of 5 nL. The protein sample is loaded to the disc by a non-contact nL-dispenser with a minimal dosage volume of 1 nL and dead volume of only 500 nL. All liquid processing steps on the disc are accomplished by centrifugal forces caused by rotation of the disc. Up to 80mum large crystals of catalase, lysozyme, proteinase K and insulin, have been produced on the disc to demonstrate the proper performance. Subsequently the crystals have been analyzed in situ in an X-ray experiment without removing them from the disc.
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