基于微流控技术的蛋白质结晶及其筛选方法的研究进展

doi:10.7503/cjcu20130691

摘要/Abstract

摘要：

微流控技术以其高通量、低消耗和集成化等优点成为蛋白质结晶微型化研究的重要手段. 本文综述了基于微流控技术的蛋白质结晶技术和方法, 主要包括微泵微阀、液滴(Droplet)、滑动芯片(SlipChip)以及液滴实验室(DropLab)等技术. 此外, 还针对当前膜蛋白在结构生物学研究中的重要地位, 综述了应用于膜蛋白结晶的微流控技术的研究进展.

关键词: 微流控学, 蛋白质结晶, 基于PDMS的微泵微阀, 液滴, 膜蛋白

Abstract:

With the advantages of high throughput, low consumption and large-scale integration, microfluidics has proven to be a promising technique for protein crystallization and screening. This article reviews the recent progress of various microfluidic techniques successfully applied in protein crystallization, including PDMS-based valve and pump, droplet, SlipChip and DropLab. In addition, due to the high importance and great challenges of membrane proteins in structural biology, we further introduce the microfluidic techniques for crystallization of membrane proteins.

Key words: Microfluidics, Protein crystallization, PDMS chip-based pump and valve, Droplet, Membrane protein

中图分类号:

O651

TrendMD:

朱丽娜, 祝莹, 方群. 基于微流控技术的蛋白质结晶及其筛选方法的研究进展. 高等学校化学学报, 2014, 35(1): 1.

ZHU Lina, ZHU Ying, FANG Qun. Recent Progress of Microfluidic Techniques for Protein Crystallization and Screening^†. Chem. J. Chinese Universities, 2014, 35(1): 1.

图/表 6

Fig.1 PDMS-based microvalve for protein crystallization under FID mode(A) Schematic illustration of PDMS microvalves under open(A1) and close(A2) states; (B) CCD image of microchip designed to implement three FID assays at three mixing ratios; (C) optical micrographs of crystals grown in the microchamber of FID chip[24].

Fig.2 PDMS peristaltic micropump and formulator chip(A)Schematic diagram of an PDMS peristaltic micropump[22]; (B) CCD image of the formulator chip[27].

Fig.3 Droplet-based microfluidic system for protein crystallization(A) Schematic diagram showing the procedures of droplet formation in a T junction channel[36]; (B) Protein crystallization with vapor diffusion method in a PDMS/glass-capillary composite microchip[35]; (C) a high throughput screening system using an on-chip integrated sampling probe and a slotted-vial array; (C1)—(C3) sample introduction process; (C4) image of a generated droplet array containing different samples in the chip[20].

Fig.4 DropLab for protein crystallization[21](A) Principle and procedure of DropLab for droplet generation; (B) setup of the DropLab system with slotted-vial array for sample presentation and microcapillaries for droplet storage; (C) protein crystallization screening result using DropLab.

Fig.5 SlipChip-based protein crystallization[46](A) Operation procedures of SlipChip; (B) crystals of the photosynthetic reaction center from Blastochloris viridis obtained with SlipChip.

Fig.6 Microfluidic system for membrane protein crystallization within lipidic mesophases(LCP)(A1) CCD image showing the mixing of bacteriorhodopsin solution(left and right chambers) with lipid monoolein(center chamber) in a microfluidic chip[63]; (A2) crystals of bacteriorhodopsin obtained in LCP; (B) membrane protein crystals grown in droplets; (B1) bacteriorhodopsin from Halobacterium salinarum; (B2) caro-tenoid-containing RC from Rhodobacter sphaeroides; (B3) caroteniodless RC from Rhodobacter sphaeroides; (B4) photosynthetic reaction center from Rhodopseudomonas viridis[65].

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