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Design of five‐layer gold nanoparticles self‐assembled in a liquid open tubular column for ultrasensitive nano‐LC‐MS/MS proteomic analysis of 80 living cells
Author(s) -
Shao Xi,
Zhang Xiangmin
Publication year - 2017
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201600463
Subject(s) - colloidal gold , nanoparticle , chromatography , chemistry , detection limit , layer (electronics) , analytical chemistry (journal) , nanotechnology , materials science
In this work, for the first time, a liquid open tubular column modified by five‐layer gold nanoparticles and linked with C 18 (GNPs@C 18 ) was designed and fabricated for nano‐LC‐MS/MS analysis of 80 living cells. Sixty nanometer gold nanoparticles were self‐assembled layer by layer on the inner wall of a 20 μm id fused‐silica capillary. C 18 was then linked on the gold nanoparticles to make the liquid open tubular column show hydrophobic character. Enough loading capacities for analysis of 80 living cells, ∼100 fmol for pk‐10 and ∼30 fmol for insulin, were obtained with the 2 m × 20 μm id five‐layer GNPs@C 18 open tubular column. The open tubular column was used in an online pretreatment and direct nano‐LC‐MS/MS analysis system to analyze 80 living HepG2 cells. In total, 650 proteins were identified in triplicate runs. The subcellular localization of the identified proteins showed that our system had no bias toward different cellular compartments. Protein copy number per cell of the identified proteins showed that the detection limit could reach 50 zmol and the abundance of the identified proteins could cover a dynamic range of 6 orders.