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Antibody-free, targeted mass-spectrometric approach for quantification of proteins at low picogram per milliliter levels in human plasma/serum

Cited 180 time in Web of Science Cited 184 time in Scopus
Authors

Shi, Tujin; Fillmore, Thomas L.; Sun, Xuefei; Zhao, Rui; Schepmoes, Athena A.; Hossain, Mahmud; Xie, Fang; Wu, Si; Kim, Jong-Seo; Jones, Nathan; Moore, Ronald J.; Pasa-Tolic, Ljiljana; Kagan, Jacob; Rodland, Karin D.; Liu, Tao; Tang, Keqi; Camp, David G., II; Smith, Richard D.; Qian, Wei-Jun

Issue Date
2012-09
Publisher
National Academy of Sciences
Citation
Proceedings of the National Academy of Sciences of the United States of America, Vol.109 No.38, pp.15395-15400
Abstract
Sensitive detection of low-abundance proteins in complex biological samples has typically been achieved by immunoassays that use antibodies specific to target proteins; however, de novo development of antibodies is associated with high costs, long development lead times, and high failure rates. To address these challenges, we developed an antibody-free strategy that involves PRISM (high-pressure, high-resolution separations coupled with intelligent selection and multiplexing) for sensitive selected reaction monitoring (SRM)-based targeted protein quantification. The strategy capitalizes on high-resolution reversed-phase liquid chromatographic separations for analyte enrichment, intelligent selection of target fractions via on-line SRM monitoring of internal standards, and fraction multiplexing before nano-liquid chromatography-SRM quantification. Application of this strategy to human plasma/serum demonstrated accurate and reproducible quantification of proteins at concentrations in the 50-100 pg/mL range, which represents a major advance in the sensitivity of targeted protein quantification without the need for specific-affinity reagents. Application to a set of clinical serum samples illustrated an excellent correlation between the results obtained from the PRISM-SRM assay and those from clinical immunoassay for the prostate-specific antigen level.
ISSN
0027-8424
URI
https://hdl.handle.net/10371/201898
DOI
https://doi.org/10.1073/pnas.1204366109
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  • College of Natural Sciences
  • School of Biological Sciences
Research Area Molecular Interactomics, Proteomics, Systems Biology, 단백체학, 분자상호작용체학, 시스템생물학

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