Enhanced Protein Digestion through the Confinement of Nanozeolite-Assembled Microchip Reactors
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- 作者：Ji Ji ; Yahong Zhang ; Xiaoqin Zhou ; Jilie Kong ; Yi Tang ; Baohong Liu
- 刊名：Analytical Chemistry
- 出版年：2008
- 出版时间：April 1, 2008
- 年：2008
- 卷：80
- 期：7
- 页码：2457 - 2463
- 全文大小：322K
- 年卷期：v.80,no.7(April 1, 2008)
- ISSN：1520-6882
文摘
An on-chip microreactor was proposed toward the acceleration of protein digestion through the constructionof a nanozeolite-assembled network. The nanozeolitemicrostructure was assembled using a layer-by-layertechnique based on poly(diallyldimethylammonium chloride) and zeolite nanocrystals. The adsorption of trypsinin the nanozeolite network was theoretically studied basedon the Langmuir adsorption isotherm model. It was foundthat the controlled trypsin-containing nanozeolite networks assembled within a microchannel could act as astationary phase with a large surface-to-volume ratio forthe highly efficient proteolysis of both proteins at lowlevels and with complex extracts. The maximum proteolytic rate of the adsorbed trypsin was measured to be350 mM min-1 
g-1, much faster than that in solution.Moreover, due the large surface-to-volume ratio andbiocompatible microenvironment provided by the nanozeolite-assembled films as well as the microfluidic confinement effect, the low-level proteins down to 16 fmol peranalysis were confidently identified using the as-preparedmicroreactor within a very short residence time coupledto matrix-assisted laser desorption-time-of-flight massspectrometry. The on-chip approach was further demonstrated in the identification of the complex extracts frommouse macrophages integrated with two-dimensionalliquid chromatography-electrospray ionization-tandem massspectrometry. This microchip reactor is promising for thedevelopment of a facile means for protein identification.
g-1, much faster than that in solution.Moreover, due the large surface-to-volume ratio andbiocompatible microenvironment provided by the nanozeolite-assembled films as well as the microfluidic confinement effect, the low-level proteins down to 16 fmol peranalysis were confidently identified using the as-preparedmicroreactor within a very short residence time coupledto matrix-assisted laser desorption-time-of-flight massspectrometry. The on-chip approach was further demonstrated in the identification of the complex extracts frommouse macrophages integrated with two-dimensionalliquid chromatography-electrospray ionization-tandem massspectrometry. This microchip reactor is promising for thedevelopment of a facile means for protein identification.