TY - GEN
T1 - Microfluidic immunoprecipitation for post-translational modified protein purification
AU - Xia, Hui
AU - Mathew, Bobby
AU - John, Tom
AU - Hegab, Hisham
AU - Feng, June
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
KW - Micro- Immunoprecipitation
KW - Microfluidic Device
KW - Polydimethylsiloxane
KW - Post-Translational Modification
UR - http://www.scopus.com/inward/record.url?scp=84876067245&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84876067245&partnerID=8YFLogxK
U2 - 10.1007/978-3-642-29305-4_80
DO - 10.1007/978-3-642-29305-4_80
M3 - Conference contribution
AN - SCOPUS:84876067245
SN - 9783642293047
T3 - IFMBE Proceedings
SP - 297
EP - 300
BT - World Congress on Medical Physics and Biomedical Engineering
T2 - World Congress on Medical Physics and Biomedical Engineering
Y2 - 26 May 2012 through 31 May 2012
ER -