Department of Physics and Optical Science and University of North Carolina at Charlotte, Charlotte, North Carolina, USA.
Department of Chemistry, University of North Carolina at Charlotte, Charlotte, North Carolina, USA.
Biopreserv Biobank. 2022 Oct;20(5):451-460. doi: 10.1089/bio.2022.0035. Epub 2022 Sep 2.
Cold-chain storage can be challenging and expensive for the transportation and storage of biologics, especially in low-resource settings. Nucleic acid nanoparticles (NANPs) are an example of new biological products that require refrigerated storage. Light-assisted drying (LAD) is a new processing technique to prepare biologics for anhydrous storage in a trehalose amorphous solid matrix at ambient temperatures. In this study, LAD was used to thermally stabilize four types of NANPs with differing structures and melting temperatures. Small volume samples (10 μL) containing NANPs were irradiated with a 1064 nm laser to speed the evaporation of water and create an amorphous trehalose preservation matrix. Samples were then stored for 1 month at 4°C or 20°C. A FLIR C655 mid-IR camera was used to record the temperature of samples during processing. The trehalose matrix was characterized using polarized light imaging (PLI) to determine if crystallization occurred during processing or storage. Damage to LAD-processed NANPs was assessed after processing and storage using gel electrophoresis. Based on the end moisture content (EMC) as a function time and the thermal histories of samples, a LAD processing time of 30 min is sufficient to achieve low EMCs for the 10 μL samples used in this study. PLI demonstrates that the trehalose matrix was resistant to crystallization during processing and after storage at 4°C and at room temperature. The native-polyacrylamide gel electrophoresis results for DNA cubes, RNA cubes, and RNA rings indicate that the main structures of these NANPs were not damaged significantly after LAD processing and being stored at 4°C or at room temperature for 1 month. These preliminary studies indicate that LAD processing can stabilize NANPs for dry-state storage at room temperature, providing an alternative to refrigerated storage for these nanomedicine products.
冷链储存对于生物制剂的运输和储存来说具有挑战性且成本高昂，尤其是在资源匮乏的环境中。核酸纳米颗粒（NANPs）是需要冷藏储存的新型生物制品的一个例子。光辅助干燥（LAD）是一种新的处理技术，用于将生物制剂在室温下无水储存于海藻糖无定形固体基质中。在这项研究中，LAD 被用于对具有不同结构和熔点的四种 NANPs 进行热稳定化处理。 含有 NANPs 的小体积样品（10 μL）用 1064nm 激光照射以加速水的蒸发并形成无定形海藻糖保存基质。然后将样品在 4°C 或 20°C 下储存 1 个月。使用 FLIR C655 中红外相机记录样品在处理过程中的温度。使用偏光成像（PLI）对海藻糖基质进行表征，以确定在处理或储存过程中是否发生结晶。使用凝胶电泳评估 LAD 处理后的 NANPs 的损伤情况。 根据最终水分含量（EMC）作为时间的函数以及样品的热历史，对于本研究中使用的 10 μL 样品，30 分钟的 LAD 处理时间足以实现低 EMC。PLI 表明，海藻糖基质在处理过程中和在 4°C 以及室温下储存时都不易结晶。DNA 立方、RNA 立方和 RNA 环的天然聚丙烯酰胺凝胶电泳结果表明，在 LAD 处理后并在 4°C 或室温下储存 1 个月后，这些 NANPs 的主要结构没有受到明显损坏。 这些初步研究表明，LAD 处理可以稳定 NANPs 用于室温下的干燥状态储存，为这些纳米医学产品提供了冷藏储存的替代方案。
