Ultrasensitive Detection of Low-Abundance Protein Biomarkers by Mass Spectrometry Signal Amplification Assay

• Published in: Analytical chemistry (Washington) Vol. 88; no. 13; pp. 6767 - 6772
• Main Authors: Du, Ruijun, Zhu, Lina, Gan, Jinrui, Wang, Yuning, Qiao, Liang, Liu, Baohong
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 05-07-2016
• Subjects: Amplification; Animals; Aptamers, Nucleotide - chemistry; Assaying; Bioassays; Biomarkers; Biomarkers - analysis; Cattle; Cell Line, Tumor; Epithelial Cell Adhesion Molecule - analysis; Epithelial Cell Adhesion Molecule - chemistry; Gold; Gold - chemistry; Limit of Detection; Mass spectrometry; Metal Nanoparticles - chemistry; Mice; Nanoparticles; Proteins; Serum - metabolism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Thrombin; Thrombin - analysis; Thrombin - chemistry
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.6b01063

A mass spectrometry signal amplification method is developed for the ultrasensitive and selective detection of low-abundance protein biomarkers by utilizing tag molecules on gold nanoparticles (AuNPs). EpCAM and thrombin as model targets are captured by specific aptamers immobilized on the AuNPs. With laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS), the mass tag molecules are detected to represent the protein biomarkers. Benefiting from the MS signal amplification, the assay can achieve a limit of detection of 100 aM. The method is further applied to detect thrombin in fetal bovine serum and EpCAM in cell lysates to demonstrate its selectivity and feasibility in complex biological samples. With the high sensitivity and specificity, the protocol shows great promise for providing a new route to single-cell analysis and early disease diagnosis.