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PharmTech.com The Real Message Behind Commercial mRNA Products April eBook 2023 Pharmaceutical Technology ® 33 multiple applications and administration methods (see Table II). By using these LNPs, it may be possible to control the delivery of the LNP and its cargo to tar- get tissues, ensuring optimal effects of the payload with minimal side-effects. LNPs that do not activate the innate immune sys- tem are available. These are especially suited for application where repeated dosing is required (e.g., protein replacement) or in applications where the RNA payload should be recognized as autologous (e.g., autoimmune disease). F u r t he r de ve lopme nt s of l i br a r ie s a nd f a m i- l ies a re bei ng processed i n t he f ie ld s of i m mu- nomodu l at ion (ac t ivat i ng or si lenci ng) a nd t a r- geted deliver y (gene-editing, protein replacement, immuno-oncology). LNP lyophilization One of the main challenges in the rollout of COVID vaccines was the requirement for cold chain trans- por t at ion a nd storage. T he development of t her- mostable RNA–cLNPs require component-specific optimization: the ionizable lipid, the LNP, the RNA, and the formulation process. Specifically, the lyo- philization process is a challenge as LNPs are t y p- ically averse to stress conditions. A proprietar y ly- ophilization process, enabling one to lyophilize a wide range of LNPs, has been developed (etherna). Drug Delivery TABLE III. Physico chemical properties of LNPs are unchanged after freeze drying. Size (nm) PDI pH Osmolality EE (%) mRNA content (µg/mL) Before FD 76.65 ± 0.75 0.14 7.52 556 95.42 87.37 Liquid control 82.31 ± 1.29 0.16 7.33 559 94.66 85.98 After FD 89.67 ± 0.43 0.22 7.37 560 91.69 86.39 -80˚C liquid 2-8˚C FD Control 0 3 x 10 7 Bioluminescence T=5m Avg Radiance [p/s/cm 2 /sr] 4h 24h 48h -80˚C liquid 2-8˚C FD 10 0 10 1 10 2 10 3 10 4 10 5 10 6 Immunogenicity T=5m Endpoint titers Total IgG D20 D35 2 x 10 7 1 x 10 7 FIGURE 7. Expression and Immunogenicity of mRNA is retained after freeze drying. In vivo expression S-Ac-7 DOG after lyo S-Ac-7 DOG fresh 0 h 4 h 24 h FIGURE 8. In vivo expression of mRNA-LNPs before and after freeze drying.