Biotinylated Au Nanoparticle-Based Artificial Antibody for Detection of Lysozyme by the Lateral Flow Immunoassay and Enzyme-Linked Immunosorbent Assay
Biotinylated Au Nanoparticle-Based Artificial Antibody for Detection of Lysozyme by the Lateral Flow Immunoassay and Enzyme-Linked Immunosorbent Assay
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基于生物素化金纳米颗粒的人工抗体，用于通过侧流免疫分析和酶联免疫吸附分析检测溶菌酶
DOI：
10.1021/acsanm.2c02268
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发表时间：
2022-08
影响因子：
5.9
通讯作者：
Aoneng Cao
中科院分区：
文献类型：
--
作者：
Wenhao Li;Tiange Gao;Chenxi Lou;Haifang Wang;Yuanfang Liu;Aoneng Cao
基于抗体的免疫分析，如侧流免疫分析（LFIA）和酶联免疫吸附测定（ELISA）是目前在许多领域广泛使用的不可缺少的分析方法，主要是由于抗体的高灵敏度和特异性。然而，单克隆抗体的高成本和它们对高温的敏感性限制了免疫测定的应用。在此之前，我们开发了一类基于金纳米颗粒（AuNP）的人工抗体，称为Goldbody。金抗体不仅具有单克隆抗体所具有的特异性，而且其稳定性也远优于单克隆抗体。为了利用金体的特异性、稳定性和易于功能化等优点，在免疫分析中取代单克隆抗体，本文合成了生物素化的抗溶菌酶金体，并成功构建了检测溶菌酶的竞争性LFIA，检测范围为17.98-226.49 ng·mL-1。同时，生物素化抗溶菌酶Goldbody可以很容易地替代商业化BA（biotinylated antibody）-ELISA试剂盒中的检测抗体用于溶菌酶的检测，与商业化BA-ELISA试剂盒相比，其检测下限为0.34 ng·mL-1，检测范围为0.89 ~ 20 ng·mL-1。此外，生物素化的抗溶菌酶金抗体在两种测定中都具有良好的热稳定性，即使在100 °C预处理后也能准确检测加标样品，证明了金抗体作为免疫测定中单克隆抗体的良好替代品的高潜力。
Antibody-based immunoassays such as the lateral flow immunoassay (LFIA) and enzyme-linked immunosorbent assay (ELISA) are currently indispensable analytical methods widely used in many fields, mainly due to the high sensitivity and specificity of antibodies. However, the high cost of monoclonal antibodies and their susceptibility to high temperature limit the application of immunoassays. Previously, we developed a class of gold nanoparticle (AuNP)-based artificial antibodies, called Goldbody. Goldbodies not only bind antigens as specifically as monoclonal antibodies do but also have far better stability than monoclonal antibodies. To take advantage of the excellent specificity, stability, and easy functionalization of Goldbodies and use them for the substitution of monoclonal antibodies in immunoassays, herein, we synthesize a biotinylated anti-lysozyme Goldbody and successfully construct a competitive LFIA for the detection of lysozyme in the range of 17.98–226.49 ng·mL–1. At the same time, the biotinylated anti-lysozyme Goldbody can easily replace the detection antibody in the commercial BA (biotinylated antibody)-ELISA kit for the detection of lysozyme with a lower detection limit of 0.34 ng·mL–1and a wider detection range of 0.89–20 ng·mL–1compared with the commercial BA-ELISA kit. In addition, the biotinylated anti-lysozyme Goldbody has good thermal stability in both assays and can accurately detect spiked samples even after pretreatment at 100 °C, demonstrating the high potential of Goldbodies as a good replacement of monoclonal antibodies in immunoassays.
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影响因子：
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作者：
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