What we know about the biosynthetic mRNA so far (Christmas 2023 edition by Maria Gutschi PharmD)

from Twitter https://twitter.com/CanningPharm/status/1737837467984957514

  • it is highly codon optimized with 800+ N-1-methylpseudoU

  • it is highly resistant to RNAses & can last months+

  • frameshift to make aberrant proteins

  • only about 60% is full intact mRNA; remainder is truncated/fragmented and its role is not yet understood

  • forms adducts with the LNPs rendering it untranslatable but with unknown effects in the cell

  • a true western blot of process 2 batches have not been provided to the regulators (Specific Obligation 1)

  • it is a prodrug so that the precursor’s (ie the mRNA itself) effect on cell process are not understood

  • may form DNA/RNA hybrids

  • fluorometry overestimates the amount of RNA in the bulk drug substance (as DNA measured as well)

  • if the N-1-methylpseudoU is broken down, can these nucleotides be used in other mRNAs affecting their production and accuracy?

process related impurities

  • it has large amounts of residual DNA from the plasmid used to generate the mRNA

  • regulatory elements which are known to ferry DNA into the nucleus is present but were hidden from the regulators

  • there is a hidden gene with some homologous to spidrion silk not disclosed to the regulators

  • risks of insertional mutagenesis appear likely as well as oncological risks

  • amount of residual DNA underestimated using qPCR

  • role of small <200bp DNA fragments needs to be elucidated

  • DNA fragments etc could also be making adducts with unknown effects within the cell

  • polymerases and other chemicals from the IVT may be present in the LNPs with unknown effects

  • dsRNA appears to be the major impurity controlled by Pfizer/biontech

the purification of the mRNA drug substance

  • used DNAse1 which was not certified/verified for use by the EMA and was suboptimal but little done for 2 years

  • there were no chromatographic techniques used for purification

  • the filters for ultra- and diafiltration were reused after sterilization in NaOH

contaminants such as endotoxin are not measured accurately and are likely underestimated with possibility for serious and synergistic effects

Ok, anymore stuff you can add?

Is this soup in these LNPs capable of making a true spike protein? Does it fulfill its primary purpose?