A Novel Electrochemiluminescent Immunoassay Based on Target Transformation Assisted with Catalyzed Hairpin Assembly Amplification for the Ultrasensitive Bioassay
| Autor: | Ruo Yuan, Tian Li, Pu Liao, Wen-Bin Liang, Yafang Wan, Hu-Xin Lei, Chang-Chun Niu |
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| Rok vydání: | 2019 |
| Předmět: |
Tris
Materials science Metallocenes chemistry.chemical_element 02 engineering and technology 01 natural sciences Catalysis Ruthenium chemistry.chemical_compound Limit of Detection medicine Humans General Materials Science Ferrous Compounds Immunoassay Detection limit medicine.diagnostic_test Oligonucleotide 010401 analytical chemistry Nucleic Acid Hybridization DNA Electrochemical Techniques 021001 nanoscience & nanotechnology Combinatorial chemistry 0104 chemical sciences chemistry Linear range Ferrocene Biological Assay 0210 nano-technology Biosensor Cysteine-Rich Protein 61 |
| Zdroj: | ACS Applied Materials & Interfaces. 11:31427-31433 |
| ISSN: | 1944-8252 1944-8244 |
| Popis: | In this work, we constructed a novel electrochemiluminescent (ECL) strategy based on sandwich immunoassay-induced target transformation assisted with catalyzed hairpin assembly (CHA) amplification for ultrasensitive bioassay with cysteine-rich protein 61 (CCN1) as a model. First, the target CCN1 could be equally transformed into the specific oligonucleotide (initiator I) labeled on the detection antibody based on the specific sandwich immunoassay. In addition, the initiator I triggered an efficient nonenzymatic CHA amplification in the presence of ferrocene-labeled hairpin 1 (Fc-H1) and hairpin 2 (H2) to produce massive hybrids (Fc-H1-H2) containing a sticky end labeled with ferrocene. Finally, Fc-H1-H2 could be immobilized on the capture probe single-stranded DNA (ssDNA)-modified electrode through the hybridization between the sticky end of Fc-H1-H2 and ssDNA, and a significantly quenched ECL signal could be obtained due to the efficient quench effect between ferrocene and the ECL indicator, ruthenium(II) tris(4,4'-dicarboxylicacid-2,2'-bipyridyl) [Ru(dcbpy)32+], immobilized on the surface of the electrode, which was related to the concentration of target CCN1. As expected, the proposed ECL biosensor exhibited a relatively low detection limit of 3.9 fg/mL in a linear range from 10 fg/mL to 100 ng/mL. This ECL strategy inspired the clinical examination of the biomarker CCN1, providing potential application in early diagnosis and malignant monitoring of cancer. |
| Databáze: | OpenAIRE |
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