Rapid Electrophoretic Assay of mRNA Vaccine Quality Attributes

This project goal is to develop a method to rapidly assess critical quality attributes (CQA) of mRNA lipid nanoparticles (LNPs) used in mRNA vaccines.
Categories
Assays
Data
Active Immunization Countermeasures

Industry Need

  • HPLC, used for mRNA quality control, requires significant solvent and maintenance expenses. 
  • Throughput of HPLC is limited because it requires extensive wash and conditioning steps. 
  • Additionally, gel electrophoresis methods, widely used in lab-scale testing of mRNA for LNPs, are too slow to be useful in this size range, have long run times, and require frequent replacement to maintain peak sharpness. 

Approach

Carnegie Mellon aims to develop a means to rapidly assess CQAs of mRNA LNPs used in mRNA-based vaccines through capillary electrophoresis (CE). The method is a form of capillary electrophoresis (CE), a widely accepted, miniaturizable, and easily deployed analytical technique, with several commercial platforms available for use in biomanufacturing facilities and clinical diagnostic labs. CE uses a rapid, surfactant-based form of electrophoretic DNA and RNA separation that characterizes size resolution appropriate for mRNA used as vaccine feedstocks.  


Carnegie Mellon University will present: 

  1. A rapid means to characterize mRNA quality prior to LNP encapsulation 
  2. A rapid means to characterize mRNA quality following release from LNPs  
  3. A combination method that quickly measures LNP surface charge and mRNA payload quality – two CQAs linked to vaccine effectiveness. This method could be used to assay mRNA quality following their synthesis by in vitro transcription, to study the impact of shelf life and temperature history on the physical properties of mRNA LNPs, and even to verify the quality of mRNA vaccines at point-of-use.  

Impacts

The project will demonstrate a transformative means to rapidly and accurately characterize nanoparticles used in COVID-19 vaccines.

Since electrophoresis instrumentation is relatively cheap and easy to use, the method established could be deployed in biomanufacturing and routine testing of stocks at point-of-use to ensure product safety.

Value Statement/Outcomes

Adopting rapid capillary electrophoresis (CE)-based assays for mRNA LNP analysis reduces reliance on complex, high-cost workflows by using commercially available CE platforms, minimizing consumables and labor expenses. With an 8-minute runtime per sample, these assays accelerate quality verification and batch release, supporting high-throughput manufacturing. Robust control of critical quality attributes—including mRNA dosage and integrity—ensures consistent, multivalent vaccine formulations from production to point-of-use, optimizing resources and timelines.

Outputs/Deliverables

WMB-CE assay was demonstrated to give a quantitative measure of mRNA dosage in multivalent vaccines, with an 8-minute runtime on a commercial CE instrument. Further, LNP materials can be directly injected into the CE column, without modification or LNP lysis, to give mRNA dosage. Finally, the method was able to track mRNA damage by thermal degradation, enzymatic degradation, and freeze-thaw cycling.

Posters

Schneider, J., Presenter, ARP-07 Rapid Electrophoretic Assay of mRNA Vaccine Quality Attributes, NIIMBL National Meeting, Washington, D.C., July 28, 2022.

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Project Lead

Carnegie Mellon University

Carnegie Mellon University