Measurement of mRNA therapeutics: method development and validation challenges

Area of Science:

• Biopharmaceutical Development
• Analytical Chemistry
• Molecular Biology

Background:

• Chemically modified mRNA therapeutics are increasingly prevalent in drug development pipelines for diverse disease indications.
• The routine assessment of these complex analytes presents unique challenges for the pharmaceutical industry and contract research organizations (CROs).

Purpose of the Study:

• To outline the challenges associated with the regulated bioanalysis of modified mRNA therapeutics.
• To compare and contrast the two primary analytical methodologies employed for this purpose: quantitative reverse transcription PCR (RT-qPCR) and branched DNA (bDNA) assays.

Main Methods:

• Comparative analysis of quantitative reverse transcription PCR (RT-qPCR) for modified mRNA quantification.
• Comparative analysis of branched DNA (bDNA) assays for modified mRNA quantification.
• Discussion of strategies for overcoming common challenges in regulated bioanalysis of mRNA.

Main Results:

• Both RT-qPCR and bDNA assays present distinct advantages and limitations for the bioanalysis of modified mRNA therapeutics.
• Specific strategies are required to address challenges such as sensitivity, specificity, and matrix effects inherent in these analytical approaches.

Conclusions:

• The selection of an appropriate analytical method (RT-qPCR or bDNA) depends on the specific requirements of the modified mRNA therapeutic and the bioanalytical study.
• Robust bioanalytical strategies are crucial for the successful and regulated development of mRNA-based medicines.