Efficient sample proteolysis based on a microchip containing a glass fiber core with immobilized trypsin

被引:7
作者
Bao, Huimin [1 ]
Liu, Shuang [2 ]
Zhang, Luyan [1 ]
Chen, Gang [1 ]
机构
[1] Fudan Univ, Sch Pharm, Dept Chem, Shanghai 200032, Peoples R China
[2] State Ocean Adm, N China Sea Environm Monitoring Ctr, Qingdao 266033, Peoples R China
关键词
Microchip; Trypsin; Immobilization; Oxidized cellulose; Proteolysis; CAPILLARY ELECTROPHORESIS; PROTEIN DIGESTION; CHIP; IDENTIFICATION; SYSTEMS; REACTOR; CHANNEL; LAYER;
D O I
10.1007/s00604-012-0896-0
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Trypsin was immobilized on cellulose-coated glass fibers via a condensation reaction between the aldehyde groups of the oxidized cellulose and the primary amino groups of trypsin. A piece of the modified fiber was inserted into the main channel of a poly(methyl methacrylate) microchip to form a microfluidic proteolytic bioreactor. Scanning electron microscopy of the cross section of the fiber revealed a rough film on the surface of the fiber glass. The performance of the bioreactor was demonstrated by the tryptic digestion of hemoglobin and cytochrome c, where the time for digestion was reduced to < 10 s. The digests were identified by MALDI-TOF-MS to obtain peptide mass fingerprint spectra. The results indicated that the digestion in the microfluidic bioreactor is comparable to that of a 12-h solution tryptic digest and thus provides a promising platform for the high throughput identification of proteins.
引用
收藏
页码:291 / 297
页数:7
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