The vax is ~4200bp.
This is approximately 1 error in every vax molecule and you get injected with 40T.
This is a result of m1pU which is a low fidelity base. This low fidelity also impacts the next step of tRNA hybridization in translation.
This is compounding error.
Pfizer has mostly sold this base replacement as improving the magnitude and durability of expression.
As with many things in biology, when you optimize for the magnitude of expression, you sacrifice fidelity.
Many think of variants as from the virus, not the vax
The virus has an ExoN gene to error correct the polymerase errors. These error correcting proteins don’t like low fidelity bases like m1pU
In order maximize expression of m1pU they need sloppy enzymes
Add the transcriptional and translational compounded error and you have a combinatorial biochemistry problem on every injection.
This is why there is raw sequence data for vax lots in NCBI.
There is no peptide sequence of one of these mRNA libraries in NCBI.
Just smears on gels.
This is a highly variable prodrug.
The final drug has never been characterized.
There are 3D structures of protein translated from mRNA transcribed from a DNA construct without m1pU. Not relevant. No m1pU.
There are also smeary antibody stained western blots.
Antibody stains are biased toward error free proteins. They don’t tag the mutated ones.
Need to stain everything.
Cell free in vitro TNT to understand glycosylation or PNGase F for removing in vivo glycosylation. More ?s.
In summary, no mRNA injection should ever escape lot to lot sequence QC that is sensitive enough to find parts per thousand error. 40T molecules injected means small % error = billions of contaminants.
Protein sequencing of the final drug should be required.
This is why there is NO lot to lot raw sequence data in NCBI.^^^^
To simplify, the error measured by Chen et al, suggests 1 error in every mRNA molecule. Poisson would imply some molecules have 2 and 3 errors and many have zero.
Now imagine you inject 40 Trillion molecules where each one is different.
The combinatorics are mind blowing.
The good news is that folk at BASE are starting to look at this problem with direct RNA sequencing with Oxford Nanopore (ONT). But the m1pUs look foreign to the ONT platform and get called as both a C and a T. It's unlikely this will have the accuracy to pick up 1:1K heteroplasmy
The bad news is, this should have been done and made public before injecting 1B people. Its pretty bleeding edge so I can see how it was overlooked but at the minimum ILMN sequencing could have measured the heteroplasmies. Probably need UMIs to discount the cDNA syn error.
But now that the camel has its nose in the tent and has feasted on the money machine.. It will be back for more.
For those doubting my assessment of the mfg process, BASE spells it out here. Maybe my purity expectations are too high but I'd be anlot less scrupulous if Pfizer/Moderna put any raw sequence live for lot to lot QC. Zero for the vax. Millions for the virus.
This preprint monitors the translation fidelity in PseudoU and m1PseudoU.
If anyone has spare cycles, it would be helpful to calculate the Amino Acid edit distance of The vax Amino Acids to human amino acids in all 6 reading frames. This would give us a framework of the autoimmune risk of these synthesis errors.
The EMA leak has a lot to say about RNA integrity loss during manufacturing scale up.
If I had to guess,
Nucleotide purity probably played a role. It’s hard to scale up the synthesis of modified nucleotides and the polymerases don’t like to incorporate them. They stall.
Particularly at long tracks of U.
PolyU incorporation it pure N1 methyl pseudoU could create polymerase stall points and truncated mRNA.
But RNA integrity numbers also include mRNAs that are too long so one can’t assume it’s only truncated mRNA.
Hanna et al found it in Breast milk 5 days later
Krauson found it in heart milk issue 30 days later
Castruita found it in plasma 28 days later
Gonzalez found it in Placenta 10 days later.
Just keep in mind, the PCR data in the cell lines is solid as we’re using assays we know work and sequencing shows it’s there.
The integrations need replication and validation.
We have 2+ unique reads for each event. But we don’t have both junctions covered.
Need to sequence deeper and with longer reads and it’s need to be replicated by others.
We did not find these in the unvaxxed control OvCar3 samples.
The odds of this being chimeric ligation at the same base in the Illumina library prep is 1 in 3 billion.
There is a possibility it is chromothripsis where the genome is shattered in cancer and the chimeric reads are extrachromosomal.
Hence longer reads are required to pin this down.
We sequenced vax treated OvCar3 cell lines that were washed and passaged such that the vaccine was diluted out and only cells with DNA inside are present.
Our collaborators who did this work also stained for spike expression.
60% of cells are positive for spike with IHC
We qPCRd these cells for spike,SV40 and Ori in bulk.
Not single cell measurement.
Positive with CTs similar to human RNAP.
We performed whole genome sequencing.
The entire vax plasmid can be reassembled at 3,000X coverage (Top Track).
This is at 30X coverage of the human genome so 100x higher coverage for the plasmid than the human genome?
As a control we also sequenced the vax alone.
It’s at 44,000X coverage.
You can see the left side of the IGV view has no SNPs. That’s the vector.
The spike has SNPs
Anyone versed in Aspergillus must know about AF36 and Afla-Guard.
This is a point mutation that knocks out the aflatoxin producing genes and is used on crops to outcompete wildtype mycotoxin producing Aspergillus flavus.
So there is an entire Agriculture industry built around man modified biocontrol agents that are used to occupy a niche and outcompete wild strains since 2004.
So it’s very bizarre to have an Aspergillus ‘expert’ make such counter factual points.