Label-free and amplified electrogenerated chemiluminescence biosensing for the detection of thymine DNA glycosylase activity using DNA-functionalized gold nanoparticles triggered hybridization chain reaction
| Autor: | Yuecheng Zhang, Yingying Wei, Wanqiao Bai, Lin Bao, Yan Li |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Base pair
Metal Nanoparticles Breast Neoplasms Biosensing Techniques 02 engineering and technology 01 natural sciences Biochemistry Analytical Chemistry law.invention chemistry.chemical_compound law Humans Environmental Chemistry AP site Spectroscopy Chemiluminescence Chemistry 010401 analytical chemistry DNA Electrochemical Techniques 021001 nanoscience & nanotechnology Combinatorial chemistry Thymine DNA Glycosylase 0104 chemical sciences Thymine Colloidal gold Luminescent Measurements Female Gold Thymine-DNA glycosylase 0210 nano-technology Biosensor |
| Zdroj: | Analytica Chimica Acta. 1061:101-109 |
| ISSN: | 0003-2670 |
| Popis: | Effective detection of thymine DNA glycosylase (TDG) activity is extremely crucial and urgent for epigenetic research. Herein, a novel label-free electrogenerated chemiluminescence (ECL) biosensing method was developed for the detection of TDG activity using DNA-functionalized gold nanoparticles (DNA-AuNPs) triggered hybridization chain reaction (HCR). In this assay, the thiol modified hairpin probe DNA (hp-DNA) with 5′ overhangs and one mismatched base pair of guanines: thymine (G: T) in the stem part was boned onto gold electrode. TDG specifically removed T base of the G: T mismatch to produce apyrimidinic (AP) sites through the N-glycosidic bond hydrolysis. The AP site was then cleaved by the catalysis of Endonuclease IV (EnIV) to generate dsDNA containing a free 3’ end in the long sequence, which serves as a complementary sequence to hybridize with the specific sequence (ssDNA1) of DNA-AuNPs. Then, the functionalized DNA-AuNPs with initiator strands (ssDNA2) could trigger HCR to form nicked double helices DNA polymer which can embed numerous ECL indicator, Ru(phen)32+, resulting in significantly increased ECL signal. The proposed strategy combined the amplification function of DNA-AuNPs triggered HCR and the inherent high sensitivity of the ECL technique, a detection limit of 1.1 × 10−5 U/μL (0.0028 ng/mL) for TDG determination was obtained. In addition, this method was successfully applied to evaluate TDG activity in cancer cell, which provides great possibility for TDG activity assay in related clinical diagnostics. |
| Databáze: | OpenAIRE |
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