Microfluidic Immunoprecipitation for Post-Translational Modified Protein Purification
we here report an antibody functionalized microimmunoprecipitation (μIP) method used for enrich lowabundant post-translational modified (PTM) proteins.The device is a fabricated by inert,nontoxic and disposable polydimethylsiloxane (PDMS) using a silane-based chemical modification protocol,which yield an antibody-terminated PDMS surfaces.In this study,the μIP device is specifically designed for 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 μIP 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 μIP 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.
Post-Translational Modification Micro- Immunoprecipitation Polydimethylsiloxane Microfluidic Device
Hui Xia Bobby Mathew Tom John Hisham Hegab June Feng
Biomedical Engineering Department, Louisiana Tech University, Ruston, USA Engineering Department, Louisiana Tech University, Ruston, USA
国际会议
World Congress on Medical Physics and Biomedical Engineering (2012年医学物理及生物医学工程国际会议(IFMBE))
北京
英文
297-300
2012-05-26（万方平台首次上网日期，不代表论文的发表时间）