Investigations into mRNA Lipid Nanoparticles Shelf-Life Stability under Nonfrozen Conditions

分散性 信使核糖核酸 化学 生物物理学 生物化学 生物 基因 有机化学
作者
Anne-Gaëlle Reinhart,Anja Osterwald,Philippe Ringler,Y. Leiser,Matthias E. Lauer,Rainer E. Martin,Christoph Ullmer,Felix F. Schumacher,Claudia Korn,Michael Keller
出处
期刊:Molecular Pharmaceutics [American Chemical Society]
卷期号:20 (12): 6492-6503 被引量:20
标识
DOI:10.1021/acs.molpharmaceut.3c00956
摘要

mRNA LNPs can experience a decline in activity over short periods (ranging from weeks to months). As a result, they require frozen storage and transportation conditions to maintain their full functionality when utilized. Currently approved commercially available mRNA LNP vaccines also necessitate frozen storage and supply chain management. Overcoming this significant inconvenience in the future is crucial to reducing unnecessary costs and challenges associated with storage and transport. In this study, our objective was to illuminate the potential time frame for nonfrozen storage and transportation conditions of mRNA LNPs without compromising their activity. To achieve this goal, we conducted a stability assessment and an in vitro cell culture delivery study involving five mRNA LNPs. These LNPs were constructed by using a standard formulation similar to that employed in the three commercially available LNP formulations. Among these formulations, we selected five structurally diverse ionizable lipids─C12-200, CKK-E12, MC3, SM-102, and lipid 23─from the existing literature. We incorporated these lipids into a standard LNP formulation, keeping all other components identical. The LNPs, carrying mRNA payloads, were synthesized by using microfluidic mixing technology. We evaluated the shelf life stability of these LNPs over a span of 9 weeks at temperatures of 2-8, 25, and 40 °C, utilizing an array of analytical techniques. Our findings indicated minimal impact on the hydrodynamic diameter, zeta potential, encapsulation efficiency, and polydispersity of all LNPs across the various temperatures over the studied period. The RiboGreen assay analysis of LNPs showed consistent mRNA contents over several weeks at various nonfrozen storage temperatures, leading to the incorrect assumption of intact and functional LNPs. This misunderstanding was rectified by the significant differences observed in EGFP protein expression in an in vitro cell culture (using HEK293 cells) across the five LNPs. Specifically, only LNP 1 (C12-200) and LNP 4 (SM-102) exhibited high levels of EGFP expression at the start (T0), with over 90% of HEK293 cells transfected and mean fluorescence intensity (MFI) levels exceeding 1. Interestingly, LNP 1 (C12-200) maintained largely unchanged levels of in vitro activity over 11 weeks when stored at both 2-8 and 25 °C. In contrast, LNP 4 (SM-102) retained its functionality when stored at 2-8 °C over 11 weeks but experienced a gradual decline of in vitro activity when stored at room temperature over the same period. Importantly, we observed distinct LNP architectures for the five formulations through cryo-EM imaging. This highlights the necessity for a deeper comprehension of structure-activity relationships within these complex nanoparticle structures. Enhancing our understanding in this regard is vital for overcoming storage and stability limitations, ultimately facilitating the broader application of this technology beyond vaccines.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
ARIA完成签到 ,获得积分10
3秒前
科研通AI2S应助U9A采纳,获得10
4秒前
温暖小松鼠完成签到 ,获得积分10
11秒前
yy完成签到 ,获得积分10
12秒前
zeannezg完成签到 ,获得积分10
19秒前
ng完成签到 ,获得积分10
23秒前
25秒前
隐形曼青应助称心的新之采纳,获得10
27秒前
落后的怀梦完成签到 ,获得积分10
30秒前
唠叨的可燕完成签到 ,获得积分10
30秒前
超帅的又槐完成签到,获得积分10
31秒前
heroiheart'发布了新的文献求助10
32秒前
稳重乌冬面完成签到 ,获得积分10
34秒前
Silence完成签到 ,获得积分10
36秒前
40秒前
mmmaosheng完成签到,获得积分10
44秒前
大力听芹发布了新的文献求助10
45秒前
deniroming完成签到,获得积分10
45秒前
美好灵寒完成签到 ,获得积分10
48秒前
drz完成签到 ,获得积分10
48秒前
303完成签到 ,获得积分10
51秒前
李骁完成签到 ,获得积分10
54秒前
lilylian完成签到,获得积分10
57秒前
称心的新之完成签到,获得积分10
59秒前
陈晶完成签到 ,获得积分10
59秒前
wyh295352318完成签到 ,获得积分10
1分钟前
自信白凡完成签到 ,获得积分10
1分钟前
木木杉完成签到 ,获得积分10
1分钟前
Ava应助U9A采纳,获得10
1分钟前
666完成签到 ,获得积分10
1分钟前
温暖的颜演完成签到 ,获得积分10
1分钟前
既然寄了,那就开摆完成签到 ,获得积分10
1分钟前
雪飞杨完成签到 ,获得积分10
1分钟前
JD完成签到 ,获得积分10
1分钟前
heroiheart'完成签到,获得积分10
1分钟前
xkhxh完成签到 ,获得积分10
1分钟前
张先生完成签到 ,获得积分10
1分钟前
大力听芹完成签到,获得积分10
1分钟前
毛毛完成签到,获得积分10
1分钟前
Hiram完成签到,获得积分10
1分钟前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
Cognitive Neuroscience: The Biology of the Mind 1000
Technical Brochure TB 814: LPIT applications in HV gas insulated switchgear 1000
Immigrant Incorporation in East Asian Democracies 600
Nucleophilic substitution in azasydnone-modified dinitroanisoles 500
不知道标题是什么 500
A Preliminary Study on Correlation Between Independent Components of Facial Thermal Images and Subjective Assessment of Chronic Stress 500
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3968559
求助须知:如何正确求助?哪些是违规求助? 3513358
关于积分的说明 11167368
捐赠科研通 3248732
什么是DOI,文献DOI怎么找? 1794465
邀请新用户注册赠送积分活动 875065
科研通“疑难数据库(出版商)”最低求助积分说明 804664