The Application of Light-Assisted Drying to the Thermal Stabilization of Nucleic Acid Nanoparticles.

IF 1.2 4区 生物学 Q4 CELL BIOLOGY Biopreservation and Biobanking Pub Date : 2022-10-01 Epub Date: 2022-09-02 DOI:10.1089/bio.2022.0035
Phuong Anh Lam, Daniel P Furr, Allison Tran, Riley Q McKeough, Damian Beasock, Morgan Chandler, Kirill A Afonin, Susan R Trammell
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引用次数: 2

Abstract

Background: 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. Methods: 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. Results: 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. Conclusions: 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.

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光辅助干燥在核酸纳米粒子热稳定中的应用。
背景:对于生物制品的运输和储存来说,冷链储存可能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(NANP)是需要冷藏的新型生物产品的一个例子。光辅助干燥(LAD)是一种新的加工技术,用于制备在环境温度下在海藻糖无定形固体基质中无水储存的生物制品。在本研究中,LAD用于热稳定具有不同结构和熔融温度的四种类型的NANP。方法:小体积样本(10 μL)用1064 nm激光以加速水的蒸发并产生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。FLIR C655中红外相机用于记录处理过程中样品的温度。使用偏振光成像(PLI)对海藻糖基质进行表征,以确定在加工或储存过程中是否发生结晶。使用凝胶电泳在处理和储存后评估LAD处理的NANP的损伤。结果:根据最终含水量(EMC)作为函数时间和样品的热历史,LAD处理时间为30 min就足以实现10 μL样品。PLI证明海藻糖基质在加工过程中以及在4°C和室温下储存后耐结晶。DNA立方体、RNA立方体和RNA环的天然聚丙烯酰胺凝胶电泳结果表明,经过LAD处理并在4°C或室温下储存1个月后,这些NANP的主要结构没有受到显著损伤。结论:这些初步研究表明,LAD处理可以稳定纳米药物在室温下的干态储存,为这些纳米药物产品的冷藏储存提供了一种替代方案。
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来源期刊
Biopreservation and Biobanking
Biopreservation and Biobanking CELL BIOLOGY-MEDICAL LABORATORY TECHNOLOGY
CiteScore
3.30
自引率
12.50%
发文量
114
审稿时长
6-12 weeks
期刊介绍: Biopreservation and Biobanking is the first journal to provide a unifying forum for the peer-reviewed communication of recent advances in the emerging and evolving field of biospecimen procurement, processing, preservation and banking, distribution, and use. The Journal publishes a range of original articles focusing on current challenges and problems in biopreservation, and advances in methods to address these issues related to the processing of macromolecules, cells, and tissues for research. In a new section dedicated to Emerging Markets and Technologies, the Journal highlights the emergence of new markets and technologies that are either adopting or disrupting the biobank framework as they imprint on society. The solutions presented here are anticipated to help drive innovation within the biobank community. Biopreservation and Biobanking also explores the ethical, legal, and societal considerations surrounding biobanking and biorepository operation. Ideas and practical solutions relevant to improved quality, efficiency, and sustainability of repositories, and relating to their management, operation and oversight are discussed as well.
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