한빛사논문
- 조회1,626
Alfredo Quijano-Rubio1,2,8, Hsien-Wei Yeh1,8, Jooyoung Park1,6, Hansol Lee3, Robert A. Langan1,7, Scott E. Boyken1,7, Marc J. Lajoie1,7, Longxing Cao1, Cameron M. Chow1, Marcos C. Miranda1, Jimin Wi4, Hyo Jeong Hong4, Lance Stewart1, Byung-Ha Oh1,3,* & David Baker1,5,*
1Department of Biochemistry and Institute for Protein Design, University of Washington, Seattle, WA, USA. 2Department of Bioengineering, University of Washington, Seattle, WA, USA. 3Department of Biological Sciences, KAIST Institute for the Biocentury, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea. 4Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon, Republic of Korea. 5Howard Hughes Medical Institute, University of Washington, Seattle, WA, USA. 6Present address: Sana Biotechnology, Inc, Seattle, WA USA. 7Present address: Outpace Bio, Inc., Seattle, WA, USA. 8These authors contributed equally: Alfredo Quijano-Rubio, Hsien-Wei
*Correspondence David Baker or Byung-Ha Oh
Abstract
Naturally occurring protein switches have been repurposed for developing novel biosensors and reporters for cellular and clinical applications1, but the number of such switches is limited, and engineering them is often challenging as each is different. Here, we show that a very general class of protein-based biosensors can be created by inverting the flow of information through de novo designed protein switches in which binding of a peptide key triggers biological outputs of interest2. The designed sensors are modular molecular devices with a closed dark state and an open luminescent state; binding of the analyte of interest drives switching from the closed to the open state. Because the sensor is based purely on 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 demonstrate the modularity of this platform by creating biosensors that, with little optimization, 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 (HBV) antibody that achieve the sub-nanomolar sensitivity necessary to detect clinically relevant concentrations of these molecules. Given the current need for diagnostic tools for tracking COVID-193, we used the approach to design sensors of antibodies against SARS-CoV-2 protein epitopes and of the receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein. The latter, which incorporates a de novo designed RBD binder4, has a limit of detection of 15 pM and a signal over background of over 50-fold. 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.
논문정보
- Research article
- 2021년 01월 (BRIC 등록일 2021-02-01)
- 국내(교신)+국외 연구진
- 생명과학 > 분자생물학
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