Thank you. This method involves the isolation and purification of residual DNA, which may result in the loss of quantifiable DNA that should be detected and measured. Additionally, qPCR underestimates the amount of residual DNA due to its principle.
In this context, I am concerned that fragments of E. coli genomic DNA may also be encapsulated within the LNPs in the vaccine.
Thanks! In the research by Mr. McKernan et al, E. coli genomic DNA was not detected. Does this mean that the concentration is actually below the detection limit? I am currently interested in the qPCR results for E. coli genomic DNA.
Do you see that the identity for restriction mapping was removed??? Section 3.2.S.3.5.4? So you cant compare the sequence of the newer circular DNA plasmids to the original master cell bank that would have had the SV40 in it. Is that right? I might want to ask Kevin McKernan or Jonathan Gilthorpe or the like about this.
Table 3.2.S.3-3. What is the redacted non-compendial raw material?
Table 3.2.S.2.3-5 What is the acceptance criteria for residual HOST RNA, DNA, total protein and as per Geoff, endotoxin and total bioburden.
Thanks for this great Information, I was surprised why no plasmid map and the sv40 enhancer was nowhere found, because this was explicitly requested to see the sv40 inclusion as compared to the version of 2020! I guess it was omitted on purpose! Thanks for your valuable work dear Maria!
Table 3.2.S.3-3. the redacted non-compendial raw material is most likely DNase I
Table 3.2.S.2.3-5 values most likely identical to another document we have. Host Cell RNA <5% w/w, Residual Host Cell DNA <10% w/w, Residual Kanamycin <10ng/mg, Total residual Protein <10%w/w, Bioburden, i.e. Live Bacteria <1 CFU per 10ml, Endotoxin <20 EU/ml
Hi Geoff, do you have a link to the document you mentioned? this would be very important for the appeal, because if there is information publicly available, EMA has to unredact that section. Thanks!!!
Ok, I realized that your referred to the Rapporteur Rolling Review critical assessment report. Quality aspects, I know this one, but because it was 'stolen', the EMA still withholds the information
Thanks Geoff. How do you know that? The reason I ask is because I note the pH redactions. NZ documentation shows an alteration to the PBS buffer product Dec '21 to add sodium hydroxide and hydrochloric acid. Is that potentially a pH alternating change? I'm no chemist.
We know the latest date because it was mentioned in Rapporteur Rolling Review critical assessment report. Quality aspects. COVID-19 mRNA Vaccine BioNTech
Modified mRNA, encoding full length SARS-CoV-2 spike protein
EMEA/H/C/005735/RR/xxx
Written by Filip Josephson, Jean Michel RACE and Vanessa Seguin
Do you / we have the physical structure of all or any plasmics injected as part of the mRNA vaccines, or any other new toy invented in 2020. I ask in terms of Hairpin loops and linear or inverted repeats thus giving rise to palindromes. These will define how the exogenous DNA will enter and further move within the eucaryotic genome, and also further superimpositions of foregin DNA. I am now with the North Group, so contact me via the North Groups with any such data. I belive you have something from me from the North Group.
Thanks, at the moment I am only aware of the few CTD releases of Comirnaty that I published - but EMA is not publicly disclosing the documents they release under the Reg 1049/01. We hope the EMA Transparency Initiative which requests full transparency of the CTD files of Comirnaty/Biontech-Pfizer and Spikevax/Moderna will bring some light to the darkness!
I would like to see the Endotoxin Limit hidden on pages 11 and 18 because they lied about it to get EUA, later admitting they had no control.
https://geoffpain.substack.com/p/pfizer-lied-about-endotoxin-to-get
Endotoxin is required to activate Germline DNA transcription and Class Switch Recombination of the DNA contaminant.
https://geoffpain.substack.com/p/yin-yang-1-requires-endotoxin-to
Thanks, I will include your posts in the appeal against the redactions, very valuable information!
3.2.S.2.3.5.9. Host Cell DNA by qPCR
This assay for determination of host cell DNA of the circular plasmid DNA is based on the principle of the quantitative PCR. •••
I don't quite understand the interpretation here. Does this refer to the method for measuring the genomic DNA of E. coli, not the plasmid DNA?
https://www.ghr.agency/wp-content/uploads/2025/03/R_3.2.S.2.3-Control-of-Materials-Generation-of-Plasmids-AGC.pdf
Very interesting question - are there any scientists out there who can interpret these data?
Thank you. This method involves the isolation and purification of residual DNA, which may result in the loss of quantifiable DNA that should be detected and measured. Additionally, qPCR underestimates the amount of residual DNA due to its principle.
In this context, I am concerned that fragments of E. coli genomic DNA may also be encapsulated within the LNPs in the vaccine.
Yes, we know that Australia's TGA is concerned about various Bacterial contaminants and actually measured them.
https://geoffpain.substack.com/p/picograms-of-double-stranded-rna
Thanks! In the research by Mr. McKernan et al, E. coli genomic DNA was not detected. Does this mean that the concentration is actually below the detection limit? I am currently interested in the qPCR results for E. coli genomic DNA.
Due to its properties, a person familiar with BioTech says that quantification of short contaminant DNA is not possible with the qPCR method.
Fluorometer + Oxford NanoPore reagent is said to be essential.
Everything related to numbers is important, and I think it is necessary to check how the numbers that came out came out.
Do you see that the identity for restriction mapping was removed??? Section 3.2.S.3.5.4? So you cant compare the sequence of the newer circular DNA plasmids to the original master cell bank that would have had the SV40 in it. Is that right? I might want to ask Kevin McKernan or Jonathan Gilthorpe or the like about this.
Table 3.2.S.3-3. What is the redacted non-compendial raw material?
Table 3.2.S.2.3-5 What is the acceptance criteria for residual HOST RNA, DNA, total protein and as per Geoff, endotoxin and total bioburden.
Thanks
Thanks for this great Information, I was surprised why no plasmid map and the sv40 enhancer was nowhere found, because this was explicitly requested to see the sv40 inclusion as compared to the version of 2020! I guess it was omitted on purpose! Thanks for your valuable work dear Maria!
Table 3.2.S.3-3. the redacted non-compendial raw material is most likely DNase I
Table 3.2.S.2.3-5 values most likely identical to another document we have. Host Cell RNA <5% w/w, Residual Host Cell DNA <10% w/w, Residual Kanamycin <10ng/mg, Total residual Protein <10%w/w, Bioburden, i.e. Live Bacteria <1 CFU per 10ml, Endotoxin <20 EU/ml
Hi Geoff, do you have a link to the document you mentioned? this would be very important for the appeal, because if there is information publicly available, EMA has to unredact that section. Thanks!!!
Ok, I realized that your referred to the Rapporteur Rolling Review critical assessment report. Quality aspects, I know this one, but because it was 'stolen', the EMA still withholds the information
All documents known to exist can be "discovered" in Court under British Law.
Do you know the date of this report please?
It was written before 19 November 2020.
Thanks Geoff. How do you know that? The reason I ask is because I note the pH redactions. NZ documentation shows an alteration to the PBS buffer product Dec '21 to add sodium hydroxide and hydrochloric acid. Is that potentially a pH alternating change? I'm no chemist.
We know the latest date because it was mentioned in Rapporteur Rolling Review critical assessment report. Quality aspects. COVID-19 mRNA Vaccine BioNTech
Modified mRNA, encoding full length SARS-CoV-2 spike protein
EMEA/H/C/005735/RR/xxx
Written by Filip Josephson, Jean Michel RACE and Vanessa Seguin
The alteration to the Buffer without any clinical trial replaced the PBS buffer with Tromethamine (Tris).
https://geoffpain.substack.com/p/tromethamine-is-a-hazardous-substance
Altering. Apparently I'm no proof reader either.
Hi Cathy, you can edit comments by clicking on the 3 dots.
Thank you Silvia
Do you / we have the physical structure of all or any plasmics injected as part of the mRNA vaccines, or any other new toy invented in 2020. I ask in terms of Hairpin loops and linear or inverted repeats thus giving rise to palindromes. These will define how the exogenous DNA will enter and further move within the eucaryotic genome, and also further superimpositions of foregin DNA. I am now with the North Group, so contact me via the North Groups with any such data. I belive you have something from me from the North Group.
very many thanks
Stephen McGill
Thanks, at the moment I am only aware of the few CTD releases of Comirnaty that I published - but EMA is not publicly disclosing the documents they release under the Reg 1049/01. We hope the EMA Transparency Initiative which requests full transparency of the CTD files of Comirnaty/Biontech-Pfizer and Spikevax/Moderna will bring some light to the darkness!
This document is only for "manufacturing material" plasmid, not for the final product.
So, the quantitative method of residual DNA derived from plasmid DNA is not mentioned.
Thanks!!!! We have to request all the information from the plasmid DNA once it is manufactured!!!!