title: Fluorescence Immunoassay System via Enzyme-Enabled in Situ Synthesis of Fluorescent Silicon Nanoparticles.
abstract: The emergence of fluorescent nanomaterials with excellent performances has triggered the development of fluorescence analysis technique, which possesses several advantages in the research and clinical applications. However, current strategies for fluorescence immunoassay usually involve the routine fluorophore-labeled antibody and/or awkward signal generation procedure that may not be available in conventional enzyme-linked immunosorbent assay (ELISA) systems. Herein, we circumvent this problem by imparting an exquisite signal generation mechanism to commercially available alkaline phosphatase (ALP)-based ELISA platform and putting forward a conceptual fluorescent ELISA system based on an original ALP-enabled in situ synthesis of fluorescent nanomaterials. After adding target antigen, the presence of ALP labeled on antibody catalyzes the transformation of the substrate ascorbic acid 2-phosphate into ascorbic acid. Then the resultant ascorbic acid (i.e., ascorbate) interacts with amine-containing silane molecules (no fluorescence) to produce intense cyan fluorescent silicon nanoparticles. For the proof-of-concept, alpha-fetoprotein and human immunoglobulin G are chosen as the model antigen targets, and our proposed immunoassay (designated as the nanoparticles generation-based fluorescent ELISA) enables the detection with either fluorescence spectroscopy or naked-eye readout under the ultraviolet lamp. The convincing recognition mechanism and assay performance ensure fluorescent ELISA to quantitatively evaluate the alpha-fetoprotein level in serologic test and potentially apply in the clinic diagnosis of hepatocellular carcinoma.
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun, Jilin 130022, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun, Jilin 130022, China.|Department of Chemistry, University of Science and Technology of China , Hefei, Anhui 230026, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun, Jilin 130022, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Chuanxia
authors.initials: C
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun, Jilin 130022, China.|University of Chinese Academy of Sciences , Beijing 100049, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun, Jilin 130022, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , Changchun, Jilin 130022, China.|Department of Chemistry, University of Science and Technology of China , Hefei, Anhui 230026, China.
authors.forename: Xiurong
authors.initials: X
authors.lastname: Yang
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000937:Antigen-Antibody Reactions; D006528:Carcinoma, Hepatocellular / Q000000981:diagnostic imaging*; D004797:Enzyme-Linked Immunosorbent Assay; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D008113:Liver Neoplasms / Q000000981:diagnostic imaging*; D015394:Molecular Structure; D053758:Nanoparticles / Q000737:chemistry / Q000378:metabolism*; D012825:Silicon / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D000509:alpha-Fetoproteins / Q000737:chemistry / Q000276:immunology
chemical_list: D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D000509:alpha-Fetoproteins; D000469:Alkaline Phosphatase; D012825:Silicon