4.7 Article

A label-free sensitive method for membrane protein detection based on aptamer and AgNCs transfer

Journal

TALANTA
Volume 175, Issue -, Pages 470-476

Publisher

ELSEVIER
DOI: 10.1016/j.talanta.2017.07.071

Keywords

Silver nanocluster; Membrane protein; Tyrosine kinase; Aptamer; DNA

Funding

  1. National Natural Science Foundation of China [21636010, 21476266]

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Recently, membrane proteins have been considered as candidate cancer biomarkers and drug targets, due to their important roles in numerous physiological processes. Therefore, a facile, sensitive and quantitative detection of the membrane proteins is crucial for better understanding their roles in cancer cells and further validating their function in clinical research. We report a highly facile and sensitive detection method for membrane proteins on living cells in situ based on membrane protein-triggered release of cytosine (C)-rich single-stranded DNA (ssDNA) sequences, and the subsequent silver nanoclusters (AgNCs) transfer from polymer to C -rich ssDNA. The high-quantum yield and stable DNA-AgNCs allow the accurate detection of membrane proteins with facile operations and a common fluorescence spectrophotometer. The detection of protein tyrosine kinase-7 (PTK7), a membrane protein model, displays a response range from 30 pM to 2 nM with a detection limit of 12 pM. The expression of PTK7 on single Hela cell and CCRF-CEM cell was calculated to be 7.5 x 10(-19) mol and 1.8 x 10(-18) mol, respectively. Given the simple and facile operation of this method, this detection platform can be applied as a universal strategy for ultrasensitive detection of membrane protein on cell in situ.

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