适体
微尺度热泳
等温滴定量热法
表面等离子共振
指数富集配体系统进化
DNA
寡核苷酸
化学
生物素化
分子生物学
生物
纳米技术
生物化学
纳米颗粒
材料科学
基因
核糖核酸
作者
Gregory T. Pawel,Yuan Ma,Yuting Wu,Yi Lu,Ana Sol Peinetti
出处
期刊:Bio-protocol
[Bio-Protocol]
日期:2022-01-01
卷期号:12 (21)
被引量:1
标识
DOI:10.21769/bioprotoc.4548
摘要
Aptamers have been selected with strong affinity and high selectivity for a wide range of targets, as recently highlighted by the development of aptamer-based sensors that can differentiate infectious from non-infectious viruses, including human adenovirus and SARS-CoV-2. Accurate determination of the binding affinity between the DNA aptamers and their viral targets is the first step to understanding the molecular recognition of viral particles and the potential uses of aptamers in various diagnostics and therapeutic applications. Here, we describe protocols to obtain the binding curve of the DNA aptamers to SARS-CoV-2 using Enzyme-Linked Oligonucleotide Assay (ELONA) and MicroScale Thermophoresis (MST). These methods allow for the determination of the binding affinity of the aptamer to the infectious SARS-CoV-2 and the selectivity of this aptamer against the same SARS-CoV-2 that has been rendered non-infectious by UV inactivation, and other viruses. Compared to other techniques like Electrophoretic Mobility Shift Assay (EMSA), Surface Plasmon Resonance (SPR), and Isothermal Titration Calorimetry (ITC), these methods have advantages for working with larger particles like viruses and with samples that require biosafety level 2 facilities.
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