Wei Tianxiang, Du Dan, Zhu Mei-Jun, Lin Yuehe, Dai Zhihui
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.
ACS Appl Mater Interfaces. 2016 Mar;8(10):6329-35. doi: 10.1021/acsami.5b11834. Epub 2016 Mar 1.
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.
由蛋白质和磷酸铜（II）（Cu3(PO4)2）组成的蛋白质-无机纳米花最近引起了人们的关注。由于合成方法不需要有机溶剂，且具有独特的分级纳米结构，蛋白质-无机纳米花表现出增强的催化活性和稳定性，有望成为生物催化过程以及生物和生物医学领域的一种工具。在这项工作中，我们首先将酶、抗体和Cu3(PO4)2共固定到一种三合一的杂化蛋白质-无机纳米花中，使其具有双重功能：（1）抗体部分保留特异性捕获相应抗原的能力；（2）纳米花具有增强的酶活性和稳定性以产生放大信号。所制备的抗体-酶-无机纳米花首先应用于酶联免疫吸附测定，作为一种新型酶标抗体用于检测大肠杆菌O157:H7。检测限为60 CFU L(-1)，远优于商业ELISA系统。这种三合一的抗体（抗大肠杆菌O157:H7抗体）-酶（辣根过氧化物酶）-无机（Cu3(PO4)2）纳米花相对于商业酶-抗体偶联物具有一些优势。首先，它制备起来容易得多，不需要任何复杂的共价修饰。其次，由于它以大量抗体和酶的聚集体形式存在，具有相当高的捕获能力和催化活性。第三，由于其独特的分级纳米结构，与游离酶形式相比，它具有更高的酶稳定性。
