Rapid Identity and Quantity CQA Test for Multivalent mRNA Drug Product Formulations

Area of Science:

• Biotechnology
• Molecular Biology
• Vaccinology

Background:

• Messenger RNA (mRNA) has emerged as a critical platform for vaccines and therapeutics, particularly highlighted by the COVID-19 pandemic.
• Current analytical methods for characterizing mRNA critical quality attributes are often singleplex, leading to inefficiencies in labor and cost.
• There is a need for optimized, high-throughput analytical techniques for mRNA characterization.

Purpose of the Study:

• To demonstrate the feasibility of a multiplexed analytical platform, VaxArray, for mRNA analysis.
• To evaluate the VaxArray platform for efficient identity verification and concentration determination of mRNA formulations.
• To assess the analytical performance of a VaxArray mRNA assay using a model system.

Main Methods:

• Development and validation of a multiplexed VaxArray assay for mRNA analysis.
• Utilized a model system with mRNA constructs encoding influenza hemagglutinin and neuraminidase.
• Characterized analytical performance metrics including specificity, sensitivity, accuracy, and precision.
• Assayed both monovalent and bivalent mRNA formulations without prior mRNA extraction or PCR amplification.

Main Results:

• The VaxArray assay provided results in under 2 hours.
• The assay demonstrated high construct specificity, suitable for analyzing bivalent mRNA mixtures.
• Achieved vaccine-relevant sensitivity (sub-µg/mL) for influenza mRNA targets.
• Reported average accuracy of 104 ± 2% and precision of 9 ± 2%.

Conclusions:

• The VaxArray platform offers a rapid, efficient, and specific method for analyzing mRNA identity and concentration.
• This multiplexed approach overcomes limitations of singleplex assays, reducing labor and cost.
• The VaxArray assay is suitable for characterizing mRNA vaccine candidates, including multivalent formulations.