Journal
BIOSENSORS & BIOELECTRONICS
Volume 90, Issue -, Pages 494-500Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2016.10.066
Keywords
RNA methylation; N6-methyladenosine; Electrochemical immunosensor; Phos-tag-biotin; Ag@SiO2 nanocomposite
Categories
Funding
- National Natural Science Foundation of China [21375079]
- Natural Science Foundation of Shandong province, China [ZR2014BQ029]
- Project of Development of Science and Technology of Shandong Province, China [2013GZX20109]
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N6-methyladenosine (m6A), a kind of RNA methylation form and important epigenetic event, plays crucial roles in many biological progresses. Thus it is essential to quantitatively detect m6A in complicated biological samples. Herein, a simple and sensitive electrochemical method was developed for m6A detection using N6-methyladenosine-5'-triphosphate (m6ATP) as detection target molecule. In this detection strategy, anti-m6A antibody was selected as m6A recognition and capture reagent, silver nanoparticles and amine-PEG3-biotin functionalized SiO2 nanospheres (Ag@SiO2) was prepared and used as signal amplification label, and phos-tag-biotin played a vital role of bridge to link m6ATP and Ag@SiO2 through the two forms of specific interaction between phosphate group of m6ATP and phos-tag, biotin and streptavidin, respectively. Under the optimal experimental conditions, the immunosensor presented a wide linear range from 0.2 to 500 nM and a low detection limit of 0.078 nM (S/N=3). The reproducibility and specificity were acceptable. Moreover, the developed method was also validated for detect m6A content in human cell lines. Importantly, this detection strategy provides a promising immunodetection platform for ribonucleotides and deoxyribonucleotides with the advantages of simplicity, low-costing, specificity and sensitivity.
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