Introduction Lipid nanoparticles (LNPs) are the delivery system that made mRNA therapeutics and vaccines possible. Without LNPs, mRNA would be rapidly degraded in the body and unable to reach target cells. The success of COVID-19 mRNA vaccines—Comirnaty (Pfizer-BioNTech) and Spikevax (Moderna)—is built on decades of LNP research, culminating in a...

Introduction mRNA therapeutics and vaccines are complex biological products that require rigorous quality control (QC) at every stage of manufacturing. Unlike small-molecule drugs, mRNA quality cannot be assured by a single analytical test—it demands a comprehensive panel of orthogonal methods that assess identity, integrity, purity, potency, and safety. Regulatory agencies...

Introduction In vitro transcription (IVT) produces not only the desired mRNA but also a complex mixture of impurities: residual DNA template, unreacted NTPs, enzymes, short RNA fragments, double-stranded RNA (dsRNA), and salts. Each of these impurities can compromise mRNA stability, translation efficiency, and safety—particularly for therapeutic and vaccine applications. Purification...

Introduction mRNA therapeutics and vaccines require more than just a correct coding sequence. The two ends of the mRNA molecule—the 5′ cap and the 3′ poly(A) tail—are critical structural elements that determine mRNA stability, translation efficiency, and immunogenicity. Without proper capping and polyadenylation, even the most optimally designed mRNA will...

Introduction The success of mRNA vaccines against COVID-19 would not have been possible without a critical innovation: modified nucleotides. Native mRNA molecules are inherently unstable and highly immunogenic, which limits their therapeutic efficacy. The incorporation of modified nucleoside analogues—particularly pseudouridine (Ψ) and N1-methylpseudouridine (m1Ψ)—has fundamentally addressed these challenges by extending...

Introduction In vitro transcription (IVT) is the enzymatic process that produces messenger RNA (mRNA) from a DNA template. Since the global success of COVID-19 mRNA vaccines, IVT has become one of the most strategically important technologies in biopharmaceutical manufacturing. Today, mRNA is being developed for vaccine platforms, therapeutic proteins, cancer...