期刊
NATURE
卷 591, 期 7850, 页码 482-+出版社
NATURE RESEARCH
DOI: 10.1038/s41586-021-03258-z
关键词
-
资金
- HHMI
- LG Yonam Foundation
- BK21 PLUS project of Korea
- United World Antiviral Research Network (UWARN) one of the Centers Researching Emerging Infectious Diseases 'CREIDs', NIAID [1 U01 AI151698-01]
- Audacious Project at the Institute for Protein Design
- Eric and Wendy Schmidt by recommendation of the Schmidt Futures
- Washington Research Foundation
- Nordstrom Barrier Institute for Protein Design Directors Fund
- Open Philanthropy Project Improving Protein Design Fund
- 'la Caixa' Foundation [100010434, LCF/BQ/AN15/10380003]
- Air Force Office of Scientific Research [FA9550-18-1-0297]
- [1U19AG065156-01]
- National Research Foundation of Korea [4199990314463] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Protein switches have been repurposed for biosensor development by inverting the flow of information, creating modular molecular devices with a closed dark state and an open luminescent state. These sensors, based on thermodynamic coupling, require only one target binding domain for direct readout in solution, allowing for the detection of various molecules clinically, including the SARS-CoV-2 spike protein with high sensitivity and a 50-fold higher luminescence signal than background level. The modularity and sensitivity of this platform enable rapid sensor construction for a wide range of analytes, demonstrating the power of de novo protein design in creating multi-state protein systems with new functions.
Naturally occurring protein switches have been repurposed for the development of biosensors and reporters for cellular and clinical applications(1). However, the number of such switches is limited, and reengineering them is challenging. Here we show that a general class of protein-based biosensors can be created by inverting the flow of information through de novo designed protein switches in which the binding of a peptide key triggers biological outputs of interest(2). The designed sensors are modular molecular devices with a closed dark state and an open luminescent state; analyte binding drives the switch from the closed to the open state. Because the sensor is based on the thermodynamic coupling of analyte binding to sensor activation, only one target binding domain is required, which simplifies sensor design and allows direct readout in solution. We create biosensors that can sensitively detect the anti-apoptosis protein BCL-2, the IgG1 Fc domain, the HER2 receptor, and Botulinum neurotoxin B, as well as biosensors for cardiac troponin I and an anti-hepatitis B virus antibody with the high sensitivity required to detect these molecules clinically. Given the need for diagnostic tools to track the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)(3), we used the approach to design sensors for the SARS-CoV-2 spike protein and antibodies against the membrane and nucleocapsid proteins. The former, which incorporates a de novo designed spike receptor binding domain (RBD) binder(4), has a limit of detection of 15 pM and a luminescence signal 50-fold higher than the background level. The modularity and sensitivity of the platform should enable the rapid construction of sensors for a wide range of analytes, and highlights the power of de novo protein design to create multi-state protein systems with new and useful functions.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据