Department of Biochemistry, University of Washington, Seattle, WA, USA.
Institute for Protein Design, University of Washington, Seattle, WA, USA.
Nat Biotechnol. 2022 Sep;40(9):1336-1340. doi: 10.1038/s41587-022-01280-8. Epub 2022 Apr 28.
We designed a protein biosensor that uses thermodynamic coupling for sensitive and rapid detection of neutralizing antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants in serum. The biosensor is a switchable, caged luciferase-receptor-binding domain (RBD) construct that detects serum-antibody interference with the binding of virus RBD to angiotensin-converting enzyme 2 (ACE-2) as a proxy for neutralization. Our coupling approach does not require target modification and can better distinguish sample-to-sample differences in analyte binding affinity and abundance than traditional competition-based assays.
我们设计了一种蛋白质生物传感器，利用热力学偶联作用，灵敏、快速地检测血清中针对严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 变体的中和抗体。该生物传感器是一种可切换的、笼状的荧光素酶-受体结合域 (RBD) 构建体，通过检测血清抗体对病毒 RBD 与血管紧张素转化酶 2 (ACE-2) 结合的干扰，作为中和的替代指标。我们的偶联方法不需要目标修饰，并且可以比传统的基于竞争的检测方法更好地区分样品之间分析物结合亲和力和丰度的差异。
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