Institute of Quality and Safety for Agro-products, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; Key Laboratory of Grain Quality and Safety and Testing Henan Province, Zhengzhou 450002, China.
School of Life Sciences, Henan University, Kaifeng, Henan 475004, China.
Food Chem. 2024 Jul 1;445:138756. doi: 10.1016/j.foodchem.2024.138756. Epub 2024 Feb 15.
Biomimetic methods are invariably employed to synthesize hybrid organic-inorganic multilevel structure nanoflowers with self-assembly processes in aqueous solutions, which is an ideal way to meet the challenges of immobilizing antibodies or enzymes in nanomaterial based enzyme-linked immunosorbent assay (nano-ELISA). In this study, we developed protein-inorganic hybrid 3D nanoflowers composed of bovine serum albumin (BSA), horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (IgG-HRP) and copper(Ⅱ) phosphate (BSA-(IgG-HRP)-Cu(PO)) using a self-assembly biomimetic method. The preparation process avoided the use of any organic solvent and protein immobilization did not require covalent modifications. Additionally, the unique hierarchical structure enhances the thermal and storage stability of HRP. The BSA-(IgG-HRP)-Cu(PO) hybrid 3D nanoflower was then applied to a nano-ELISA platform for pyridaben detection, achieving a 50% inhibition concentration of 3.90 ng mL. The nano-ELISA achieved excellent accuracy for pyridaben detection. Such a novel BSA-(IgG-HRP)-Cu(PO) hybrid 3D nanoflower provide an excellent reagent for small molecule immunoassay.
采用仿生方法在水溶液中通过自组装过程合成具有混合有机-无机多层次结构的纳米花，这是在纳米材料酶联免疫吸附测定（nano-ELISA）中固定抗体或酶的理想方法。在这项研究中，我们开发了由牛血清白蛋白（BSA）、辣根过氧化物酶（HRP）-共轭山羊抗小鼠 IgG（IgG-HRP）和磷酸铜（BSA-（IgG-HRP）-Cu（PO））组成的蛋白质-无机杂化 3D 纳米花，使用自组装仿生方法。该制备过程避免了使用任何有机溶剂，并且蛋白质固定不需要共价修饰。此外，独特的分层结构增强了 HRP 的热稳定性和储存稳定性。然后将 BSA-（IgG-HRP）-Cu（PO）杂化 3D 纳米花应用于吡虫啉的 nano-ELISA 平台进行检测，得到 50%抑制浓度为 3.90ng/mL。nano-ELISA 对吡虫啉的检测具有优异的准确性。这种新型的 BSA-（IgG-HRP）-Cu（PO）杂化 3D 纳米花为小分子免疫分析提供了一种优异的试剂。
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