一种用于测量 RNase A 活性的高灵敏度纳米孔平台。
A highly sensitive nanopore platform for measuring RNase A activity.
机构信息
Department of Chemistry, University of Missouri, Columbia, MO 65211, USA.
Department of Chemistry, Illinois Institute of Technology, Chicago, IL, 60616, USA.
出版信息
Talanta. 2024 Aug 15;276:126276. doi: 10.1016/j.talanta.2024.126276. Epub 2024 May 22.
Ribonuclease A (RNase A) plays significant roles in several physiological and pathological conditions and can be used as a valuable diagnostic biomarker for human diseases such as myocardial infarction and cancer. Hence, it is of great importance to develop a rapid and cost-effective method for the highly sensitive detection of RNase A. The significance of RNase A assay is further enhanced by the growing attention from the biotechnology and pharmaceutical industries to develop RNA-based vaccines and drugs in large part as a result of the successful development of mRNA vaccines in the COVID-19 pandemic. Herein, we report a label-free method for the detection of RNase A by monitoring its proteolytic cleavage of an RNA substrate in a nanopore. The method is ultra-sensitive with the limit of detection reaching as low as 30 fg per milliliter. Furthermore, sensor selectivity and the effects of temperature, incubation time, metal ion, salt concentration on sensor sensitivity were also investigated.
核糖核酸酶 A（RNase A）在多种生理和病理条件下发挥着重要作用，可作为心肌梗死和癌症等人类疾病的有价值的诊断生物标志物。因此，开发一种快速且具有成本效益的方法来高度灵敏地检测 RNase A 非常重要。由于 COVID-19 大流行中成功开发了 mRNA 疫苗，生物技术和制药行业越来越关注开发基于 RNA 的疫苗和药物，这进一步凸显了 RNase A 测定的重要性。在此，我们报告了一种通过监测纳米孔中 RNA 底物的蛋白水解切割来检测 RNase A 的无标记方法。该方法具有超高的灵敏度，检测限低至 30 fg/mL。此外，还研究了传感器的选择性以及温度、孵育时间、金属离子和盐浓度对传感器灵敏度的影响。
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