title: An Improved Ultrasensitive Enzyme-Linked Immunosorbent Assay Using Hydrangea-Like Antibody-Enzyme-Inorganic Three-in-One Nanocomposites.
abstract: Protein-inorganic nanoflowers, composed of protein and copper(II) phosphate (Cu3(PO4)2), have recently grabbed people's attention. Because the synthetic method requires no organic solvent and because of the distinct hierarchical nanostructure, protein-inorganic nanoflowers display enhanced catalytic activity and stability and would be a promising tool in biocatalytical processes and biological and biomedical fields. In this work, we first coimmobilized the enzyme, antibody, and Cu3(PO4)2 into a three-in-one hybrid protein-inorganic nanoflower to enable it to possess dual functions: (1) the antibody portion retains the ability to specifically capture the corresponding antigen; (2) the nanoflower has enhanced enzymatic activity and stability to produce an amplified signal. The prepared antibody-enzyme-inorganic nanoflower was first applied in an enzyme-linked immunosorbent assay to serve as a novel enzyme-labeled antibody for Escherichia coli O157:H7 (E. coli O157:H7) determination. The detection limit is 60 CFU L(-1), which is far superior to commercial ELISA systems. The three-in-one antibody (anti-E. coli O157:H7 antibody)-enzyme (horseradish peroxidase)-inorganic (Cu3(PO4)2) nanoflower has some advantages over commercial enzyme-antibody conjugates. First, it is much easier to prepare and does not need any complex covalent modification. Second, it has fairly high capture capability and catalytic activity because it is presented as aggregates of abundant antibodies and enzymes. Third, it has enhanced enzymatic stability compared to the free form of enzyme due to the unique hierarchical nanostructure.
authors.affiliation: Jiangsu Collaborative Innovation Centre of Biomedical Functional Materials and Jiangsu Key Laboratory of Biofunctional Materials, School of Chemistry and Materials Science, Nanjing Normal University , Nanjing, 210023, P. R. China.
authors.forename: Tianxiang
authors.initials: T
authors.lastname: Wei
authors.forename: Dan
authors.initials: D
authors.lastname: Du
authors.forename: Mei-Jun
authors.initials: MJ
authors.lastname: Zhu
authors.forename: Yuehe
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Jiangsu Collaborative Innovation Centre of Biomedical Functional Materials and Jiangsu Key Laboratory of Biofunctional Materials, School of Chemistry and Materials Science, Nanjing Normal University , Nanjing, 210023, P. R. China.
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Dai
mesh_terms: D000907:Antibodies, Bacterial / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D019453:Escherichia coli O157; D053761:Nanocomposites / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry; D012680:Sensitivity and Specificity
chemical_list: D000907:Antibodies, Bacterial; D010710:Phosphates
keywords: E. coli O157:H7; ELISA; enzyme-labeled antibody; protein−inorganic nanoflower; ultrasensitive