Navid Bizmark, Satya Nayagam, Bumjun Kim, David F. Amelemah, Dawei Zhang, Sujit S. Datta, Rodney D. Priestley, Tom Colace, Jane Wang, Robert K. Prud'homme
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引用次数: 0
摘要
将信使核糖核酸（mRNA）封装在脂质纳米粒子（LNP）载体中开发出了新一代疫苗。除了 LNPs 的物理化学特性外，mRNA 在 LNPs 中的封装效率 (EE) 也是筛选疫苗组装试验的关键因素。与 mRNA 结合后信号放大的荧光染料是开发量化 EE 的检测方法的核心。然而，在检测过程中忽略时间效应会影响检测的准确性。本文报告了 Ribogreen 检测法中染料-RNA 复合物荧光强度的时间衰减动力学，并展示了非离子表面活性剂存在时如何阻碍这一动态过程。此外，还研究了这一动态过程对计算 EE 的影响。报告还介绍了准确测定动态 mRNA 负载过程所需的校正。
Ribogreen Fluorescent Assay Kinetics to Measure Ribonucleic Acid Loading into Lipid Nanoparticle Carriers
New generations of vaccines have been developed by encapsulating messenger ribonucleic acid (mRNA) in lipid nanoparticle (LNP) carriers. In addition to the physicochemical properties of LNPs, the encapsulation efficiency (EE) of mRNA in LNPs is a key factor to screen vaccine assembly assays. Fluorescent dyes with amplified signals upon binding with mRNA are at the core of developing assays to quantify EE. However, disregarding the temporal effects during the assay impacts the accuracy of the assay. Here, the kinetics of temporal decay in fluorescence intensity of dye-RNA complex—in Ribogreen assay—are reported and shown how this dynamic process can be impeded in the presence of a nonionic surfactant. Further, the impact of this dynamic process on the calculated EE is studied. The corrections needed to accurately assay dynamic mRNA loading processes are presented.
期刊介绍：
Advanced Materials Interfaces publishes top-level research on interface technologies and effects. Considering any interface formed between solids, liquids, and gases, the journal ensures an interdisciplinary blend of physics, chemistry, materials science, and life sciences. Advanced Materials Interfaces was launched in 2014 and received an Impact Factor of 4.834 in 2018.
The scope of Advanced Materials Interfaces is dedicated to interfaces and surfaces that play an essential role in virtually all materials and devices. Physics, chemistry, materials science and life sciences blend to encourage new, cross-pollinating ideas, which will drive forward our understanding of the processes at the interface.
Advanced Materials Interfaces covers all topics in interface-related research:
Oil / water separation,
Applications of nanostructured materials,
2D materials and heterostructures,
Surfaces and interfaces in organic electronic devices,
Catalysis and membranes,
Self-assembly and nanopatterned surfaces,
Composite and coating materials,
Biointerfaces for technical and medical applications.
Advanced Materials Interfaces provides a forum for topics on surface and interface science with a wide choice of formats: Reviews, Full Papers, and Communications, as well as Progress Reports and Research News.
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