Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/67885
標題: Nanoprobe-based affinity mass spectrometry for selected protein profiling in human plasma
作者: Chou, P.H.
Chen, S.H.
Liao, H.K.
Lin, P.C.
Her, G.R.
Lai, A.C.Y.
Chen, J.H.
Lin, C.C.
Chen, Y.J.
關鍵字: amyloid-p-component
nanoparticle label technology
linked-immunosorbent-assay
prostate-specific antigen
c-reactive
protein
magnetic nanoparticles
biomarker discovery
ms analysis
serum
identification
期刊/報告no:: Analytical Chemistry, Volume 77, Issue 18, Page(s) 5990-5997.
摘要: In recent decades, magnetic nanoparticles have emerged as a promising new platform in biomedical applications, particularly bioseparations. We have developed an immunoassay using antibody-conjugated magnetic nanoparticles as an efficient affinity probe to simultaneously preconcentrate and isolate targeted antigens from biological media. We combined this probe with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI MS) to profile proteins in diluted human plasma. The nanoparticles were designed to detect several disease-associated proteins and could be used directly in MALDI MS without an elution step, thereby facilitating multiple antigen screening and the characterization of antigen variants. Plasma antigens bound rapidly (similar to 10 min) to the antibody-conjugated nanoparticles, allowing the assay to be performed within 20 min. With sensitivity of detection in the femtomole range, the nanoscale immunoassay is superior to assays using microscale particles. We applied our method to comparative protein profiling of patients with gastric cancer and healthy individuals and found differential protein expression levels associated with the disease as well as individuals. Given the flexibility of manipulating functional groups on the nanoprobes, their low cost, robustness, and simplicity of the assay, our approach shows promise for targeted proteome profiling in clinical settings.
URI: http://hdl.handle.net/11455/67885
ISSN: 0003-2700
文章連結: http://dx.doi.org/10.1021/ac050655o
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