Gao Rachel Y, Riley Christine M, Toth Evan, Blair Rebecca H, Gerold Megan N, McCormick Caitlin, Taylor Amber W, Hu Tianjing, Rowlen Kathy L, Dawson Erica D
InDevR Inc., 2100 Central Ave., Suite 106, Boulder, CO 80301, USA.
Vaccines (Basel). 2022 Oct 12;10(10):1704. doi: 10.3390/vaccines10101704.
The COVID-19 pandemic highlighted mRNA as a promising platform for vaccines and therapeutics. Many of the analytical tools used to characterize the critical quality attributes of mRNA are inherently singleplex and are not necessarily optimal from a labor and cost perspective. Here, we demonstrate the feasibility of a multiplexed platform (VaxArray) for efficient identity verification and concentration determination for both monovalent and multivalent mRNA formulations. A model system comprising mRNA constructs for influenza hemagglutinin and neuraminidase was used to characterize the analytical performance metrics for a VaxArray mRNA assay. The assay presented herein had a time to result of less than 2 h, required no PCR-based amplification nor extraction of mRNA from lipid nanoparticles, and exhibited high construct specificity that enabled application to the bivalent mixture. The sensitivity for influenza hemagglutinin and neuraminidase mRNA was sub-µg/mL, which is vaccine-relevant, and the average accuracy (%recovery of a check standard) and precision were 104 ± 2% and 9 ± 2%, respectively.
新冠疫情凸显了信使核糖核酸（mRNA）作为疫苗和治疗药物的一个有前景的平台。许多用于表征mRNA关键质量属性的分析工具本质上都是单重的，从劳动力和成本角度来看不一定是最优的。在此，我们证明了一个多重平台（VaxArray）用于高效鉴定单价和多价mRNA制剂的身份及测定其浓度的可行性。使用一个包含流感血凝素和神经氨酸酶的mRNA构建体的模型系统来表征VaxArray mRNA检测的分析性能指标。本文介绍的检测方法结果得出时间少于2小时，无需基于聚合酶链反应（PCR）的扩增，也无需从脂质纳米颗粒中提取mRNA，并且具有高构建体特异性，可应用于二价混合物。对流感血凝素和神经氨酸酶mRNA的灵敏度低于微克/毫升，这与疫苗相关，平均准确度（对照标准品的回收率%）和精密度分别为104±2%和9±2%。
