Use of self assembled magnetic beads for on-chip protein digestion
Use of self assembled magnetic beads for on-chip protein digestion
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DOI：
10.1039/b504861c
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发表时间：
2005-01-01
期刊：
影响因子：
6.1
通讯作者：
Viovy, JL
中科院分区：
文献类型：
--
作者：
Slovakova, M;Minc, N;Viovy, JL
描述了在微芯片中使用接枝的胰酶磁珠进行蛋白质消化的方法。PDMS设备使用强大的磁体来产生平行于流动的磁场，该磁场具有指向芯片通道中心的强梯度。这允许形成低流体动力阻力的磁性胰酶珠塞，作为蛋白质消化的基质。这种装置代表了一种廉价的方法来制造具有合适的蛋白质孔大小的多开管状柱。对模型多肽的水解动力学研究表明，与分批系统相比，微系统获得的消化速度提高了100倍。该系统还提供了易于更换的巨大优势，因为胎圈基质很容易被冲刷掉和更换。通过对重组人生长激素在CE和MALDI-TOF MS中的消化产物的分析，证实了重组人生长激素的高效和重复性。芯片上10分钟的MS分析和原装可溶性胰酶4h的MS分析获得了相似的序列覆盖率(约44%)。
The use of grafted trypsin magnetic beads in a microchip for performing protein digestion is described. The PDMS device uses strong magnets to create a magnetic field parallel to the flow with a strong gradient pointing through the center of the chip channel. This allows for the formation of a low-hydrodynamic resistance plug of magnetic trypsin beads that serves as a matrix for protein digestion. This device represents an inexpensive way of fabricating a multi open-tubular-like column with an appropriate pore size for proteins. Kinetics studies of the hydrolysis of a model peptide show a 100-fold increase in digestion speed obtained by the microsystem when compared to a batch wise system. This system also offers the great advantage of easy replacement, as the bead matrix is easily washed out and replaced. High performance and reproducibility for digesting recombinant human growth hormone are confirmed by analysing the digest products in both CE and MALDI-TOF MS. Similar sequence coverage (of about 44%) is obtained from MS analysis of products after 10 minutes on-chip and 4 h with soluble trypsin in bulk.