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使用P NMR光谱和CRAFT测量mRNA的量化31

Gennady Khirich^1,2, Vanessa A Noreika³, Kaitlyn Doolittle Catlin¹

¹Protein Analytical Chemistry, Genentech, Inc., South San Francisco, California, USA.

Magnetic resonance in chemistry : MRC

|February 20, 2025

概括

定量31P核磁共振光谱为测量信使RNA (mRNA) 度提供了一种可靠的方法. 这种技术精确校准了mRNA灭绝系数,克服了疫苗开发传统紫外线光谱学的局限性.

科学领域:

生物化学生物化学
分析化学分析化学
分子生物学分子生物学

背景情况:

使者RNA (mRNA) 是疫苗开发中的关键治疗药物,需要精确的量化.
传统的紫外线光谱用于mRNA测量是易受溶液条件变化的.
精确校准mRNA灭绝系数对于可靠的度确定至关重要.

研究的目的:

为准确的mRNA度测量建立定量31PNMR光谱法.
解决紫外线光谱在多种化学环境中量化mRNA的局限性.
为了能够精确校准mRNA灭绝系数用于研究和制造.

主要方法:

利用定量 31P NMR 光谱作为一个直角的方法来定量 mRNA.
开发了一种对mRNA样本的定量处理,该样本具有多个多基化序列.
通过循环组合研究,通过广泛的mRNA信号研究频率域集成的局限性.
采用时间域CRAFT量化,比集成更高的精度.

主要成果:

开发的31P核磁共振方法在测量mRNA度中引入了≤4%的偏差.
广泛的mRNA共振的频域集成存在限制和主观性.
使用31P NMR进行时间域CRAFT量化证明优于整合.
实现了精确的总mRNA量化,从而实现了精确的灭绝系数校准.

结论:

定量31PNMR光谱学为总mRNA量化提供了准确可靠的方法.
这种技术克服了紫外线光谱对mRNA测量的局限性.
精确校准mRNA灭绝系数现在是可行的,支持疫苗开发和制造.

关键词:

31P qNMR 是一个31P qNMR.工艺品 (CRAFT) 是一种手工艺品.这是NMR的NMR.紫外线光谱法紫外线光谱法它们是mRNARNA.核酸中的核酸.轮式的轮式的比赛.时间域分析时间域分析

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