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Microfluidic Immunoprecipitation for Post-Translational Modified Protein Purification
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作者 XIA Hu Bobby Mathew +2 位作者 Tom John Hisham Hegab June Feng 《Chinese Journal of Biomedical Engineering(English Edition)》 2012年第2期80-86,共7页
In this paper, we report an antibody functionalized microimmunopreci- pitation (IX IP) method used for enrich lowabundant post-translational modified (PT~ proteins. The device is fabricated by inert, nontoxic and d... In this paper, we report an antibody functionalized microimmunopreci- pitation (IX IP) method used for enrich lowabundant post-translational modified (PT~ proteins. The device is fabricated by inert, nontoxic and disposable polydimethylsiloxane (PDMS) using a silane-based chemical modification protocol, which yield antibody- terminated PDMS surfaces. In this study, the IX IP device is specifically designed for the purification of carbonylated protein, a representative example here to illustrate the potential applications for any other PTMs, which could be immuno-tagged by specific antibodies. The test model in vitro oxidized bovine serum albumin (BSA) was first derivitized by dinitrophenylhydrazide (DNPH) and then captured by the anti-DNP immobilized on this Ix lP device. The surface functional group mapping was systematically analyzed and validated by fluorescence microscopy. Quantitative study of DNP-derivatized carbonylated protein capture recovery and elution efficiency of the device was also studied. We also envision that this proteome enrichment Ix IP device can be assembled with other lab-on-a-chip components, such as microelectrophoresis or micro-chromatographic devices for follow-up protein analysis. This selective enrichment of modified proteins greatly facilitates the study of low abundant protein biomarkers discovery. 展开更多
关键词 post-translational modification microimmunoprecipitation polydi-methylsiloxane microiluidic device
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