Protein sensors combining both on-and-off model for antibody homogeneous assay.
Protein sensors combining both on-and-off model for antibody homogeneous assay.
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结合开关模型的蛋白质传感器用于抗体均质测定
DOI：
10.1016/j.bios.2022.114226
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发表时间：
2022-08-01
影响因子：
12.6
通讯作者：
Hu JL
中科院分区：
文献类型：
--
作者：
Li J;Wang JL;Zhang WL;Tu Z;Cai XF;Wang YW;Gan CY;Deng HJ;Cui J;Shu ZC;Long QX;Chen J;Tang N;Hu X;Huang AL;Hu JL
基于变构酶响应靶标结合的蛋白质传感器具有快速变化的酶活性，是进行均相分析的潜在工具。然而，在它们的建设中，很难达到高信噪比(S/N)。高的S/N是区分信号和背景的关键，这一现象可能在不同个体的血清样本中存在很大差异。在此，我们基于模块化的荧光素酶NanoLuc，设计了一种新型的生物传感器--NanoSwitch。该传感器无需洗涤步骤，即可在45分钟内直接检测1例μL血清中的抗体。在FLAG和HA抗体的检测中，纳米开关对S/N比分别为33倍和42倍的抗体产生反应。此外，我们还构建了检测SARS-CoV-2特异性抗体的NanoSwitch，检测结果显示血清样本中的S/N高达200多倍。通过一种新的工作模式实现了高S/N，该工作模式将传感器与人血清白蛋白的关闭和与特定抗体的打开相结合。此外，我们还构建了用于检测丙型肝炎病毒核心蛋白抗体和人类免疫缺陷病毒gp41抗体的纳米开关。有趣的是，这些传感器在临床样本的分析中表现出高S/N和良好的性能；这在一定程度上归因于断断续续模式的结合。综上所述，我们提供了一种新型的蛋白质传感器和一种工作模型，可以潜在地指导新的传感器设计，使其具有更好的性能。
Protein sensors based on allosteric enzymes responding to target binding with rapid changes in enzymatic activity are potential tools for homogeneous assays. However, a high signal-to-noise ratio (S/N) is difficult to achieve in their construction. A high S/N is critical to discriminate signals from the background, a phenomenon that might largely vary among serum samples from different individuals. Herein, based on the modularized luciferase NanoLuc, we designed a novel biosensor called NanoSwitch. This sensor allows direct detection of antibodies in 1 μl serum in 45 min without washing steps. In the detection of Flag and HA antibodies, NanoSwitches respond to antibodies with S/N ratios of 33-fold and 42-fold, respectively. Further, we constructed a NanoSwitch for detecting SARS-CoV-2-specific antibodies, which showed over 200-fold S/N in serum samples. High S/N was achieved by a new working model, combining the turn-off of the sensor with human serum albumin and turn-on with a specific antibody. Also, we constructed NanoSwitches for detecting antibodies against the core protein of hepatitis C virus (HCV) and gp41 of the human immunodeficiency virus (HIV). Interestingly, these sensors demonstrated a high S/N and good performance in the assays of clinical samples; this was partly attributed to the combination of off-and-on models. In summary, we provide a novel type of protein sensor and a working model that potentially guides new sensor design with better performance.
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DOI：
10.3390/s21030795
发表时间：
2021-01-25
期刊：
Sensors (Basel, Switzerland)
影响因子：
--
作者：
Kim H;Ju J;Lee HN;Chun H;Seong J
通讯作者：
Seong J
影响因子：
3.7
作者：
Engler, Carola;Kandzia, Romy;Marillonnet, Sylvestre
通讯作者：
Marillonnet, Sylvestre
影响因子：
2.9
作者：
Isazadeh, Mohsen;Amandadi, Mojdeh;Hosseinkhani, Saman
通讯作者：
Hosseinkhani, Saman
影响因子：
82.9
作者：
Long, Quan-Xin;Liu, Bai-Zhong;Huang, Ai-Long
通讯作者：
Huang, Ai-Long
影响因子：
8.9
作者：
Arts R;Ludwig SKJ;van Gerven BCB;Estirado EM;Milroy LG;Merkx M
通讯作者：
Merkx M