CLDN6 CARVac manufacturing
RNA, liposomes and RNA-LPX were manufactured under good manufacturing practice conditions. The CLDN6 CARVac RNA is a highly purified, non-nucleoside-modified, uridine-containing RNA.The single-stranded, 5′-capped mRNA was produced by in vitro transcription from the corresponding DNA template encoding CLDN6. The general structure of the CLDN6 RNA consists of 5′-cap, 5′-and 3′-untranslated regions, CLDN6 coding sequence and poly(A)-tail. In addition to the coding sequence, the RNA comprises sequence elements to enhance RNA stability and translational efficiency of the encoded antigen that have been optimized during development. The RNA backbone is optimized for translation of the coding sequence in human dendritic cellsLiposomes with net cationic charges were used to complex the RNAs to form RNA-LPX. The cationic liposomes were manufactured using an adopted proprietary protocol based on the ethanol injection technique from the cationic synthetic lipid (R)-N,N,N trimethyl-2-3-dioleyloxy-1-propanaminium chloride (R-DOTMA) (Merck and Cie) and the phospholipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine phospholipid (DOPE) (Corden Pharma)The injectable RNA-LPX drug products were prepared in a dedicated pharmacy by incubating the individual concentrated RNA drug products with an isotonic NaCl solution (0.9%) (Fresenius Kabi) and cationic liposomes according to a proprietary protocol