Development of a Universal Multiplex Assay System fot High-Throughput Biological Applications
EMSL Project ID
40991
Abstract
Recent advances in genomics, proteomics, and metabolomics make the 'omics' technologies powerful discovery-based tools for identifying candidate biomarkers for human diseases; however, it has not been successful so far to establish new biomarkers for clinical practice by utilizing these technologies. The main bottleneck lies in the lack of effective tools for high-throughput validation. To overcome this bottleneck I propose to develop a novel 'reagent-free' mass spectrometry-based universal multiplex assay system that will provide high throughput quantitative measurements for hundreds of low-abundance protein and metabolite analytes, independent of antibody-based reagents. The goal for this technology platform is to achieve a profound advance over current MS-platforms by providing >1000-fold enhancement in analyte signal intensities, sufficient for detecting low-abundance species, and >5000-fold improvement in resolving power for extremely high specificity detection. These advances will be achieved by developing and integrating 1) a novel subambient pressure ionization source with nanoelectrospray array, 2) advanced ion-funnel interfaces, 3) novel multi-stage gas-phase ion mobility technology (differential mobility analyzer coupled to field asymmetric ion mobility spectrometry) for separating and selecting analytes of interest, and (4) a new triple-stage pentaquardrupole (QqQqQ) mass spectrometer for further isolating as well as detecting ions.
Project Details
Start Date
2010-11-02
End Date
2013-11-17
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
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Duan J, VK Kodali, MJ Gaffrey, J Guo, RK Chu, DG Camp, II, RD Smith, BD Thrall, and W Qian. 2015. "Quantitative Profiling of Protein S-Glutathionylation Reveals Redox-Dependent Regulation of Macrophage Function During Nanoparticle-Induced Oxidative Stress." ACS Nano Epub ahead of print:, doi:10.1021/acsnano.5b05524
Kim JS, ME Monroe, DG Camp, II, RD Smith, and W Qian. 2013. "In-Source Fragmentation and the Sources of Partially Tryptic Peptides in Shotgun Proteomics ." Journal of Proteome Research 12(2):910-916. doi:10.1021/pr300955f.Epub 2013 January 16.
Shi T, and W Qian. 2013. "Antibody-free PRISM-SRM for multiplexed protein quantification: Is this the new competition for immunoassays in bioanalysis?" Bioanalysis 5(3):267-269. doi:10.4155/bio.12.336
Shi T, D Su, T Liu, K Tang, DG Camp, II, W Qian, and RD Smith. 2012. "Advancing the sensitivity of selected reaction monitoring-based targeted quantitative proteomics." Proteomics 12(8):1074-1092. doi:10.1002/pmic.201100436
Shi T, J Zhou, MA Gritsenko, M Hossain, DG Camp, II, RD Smith, and W Qian. 2012. "IgY14 and SuperMix immunoaffinity separations coupled with liquid chromatography-mass spectrometry for human plasma proteomic biomarker discovery." Methods 56(2):246–253. doi:10.1016/j.ymeth.2011.09.001
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Sigdel TK, Y Gao, J He, A Wang, CD Nicora, TL Fillmore, T Shi, BJM Webb-Robertson, RD Smith, W Qian, O Salvatierra, DG Camp, II, and M Sarwal. 2016. "Mining the Human Urine Proteome for Monitoring Renal Transplant Injury ." Kidney International 89(6):1244-1252. doi:10. 1016/j. kint. 2015. 12. 049
Su D, AK Shukla, B Chen, JS Kim, ES Nakayasu, Y Qu, UK Aryal, KK Weitz, TRW Clauss, ME Monroe, DG Camp, II, DJ Bigelow, RD Smith, RN Kulkarni, and W Qian. 2013. "Quantitative Site-specific Reactivity Profiling of S-Nitrosylation in Mouse Skeletal Muscle Using Cysteinyl Peptide Enrichment Coupled with Mass Spectrometry." Free Radical Biology & Medicine 57:68-78. doi:10.1016/j.freeradbiomed.2012.12.010
Su D, MJ Gaffrey, J Guo, KE Hatchell, RK Chu, TRW Clauss, JT Aldrich, S Wu, SO Purvine, DG Camp, II, RD Smith, BD Thrall, and W Qian. 2013. "Proteomic Identification and Quantification of S-glutathionylation in Mouse Macrophages Using Resin-Assisted Thiol-affinity Enrichment and Isobaric Labeling." Free Radical Biology & Medicine. doi:10.1016/j.freeradbiomed.2013.12.004
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