Process 3: Pfizer's change to Tris buffer

Pfizer gamed the FDA on its change from PBS to Tris plus a patent sheds light on the seriousness of particle size and "stable folded RNA" with the Pfizer's purple-topped vials

How it started

In the October 29, 2021 revision, FDA authorized: 1) the use of Pfizer-BioNTech COVID-19 Vaccine (Original monovalent) for children 5 through 11 years of age; and 2) a manufacturing change to include an additional formulation of the Pfizer-BioNTech COVID-19 Vaccine (Original monovalent) that uses tromethamine (Tris) buffer instead of phosphate buffered saline (PBS) used in the originally authorized Pfizer-BioNTech COVID-19 Vaccine (Original monovalent). The formulation of the Pfizer-BioNTech COVID-19 Vaccine (Original monovalent) that uses Tris buffer was authorized in two presentations:

What IS this manufacturing change?

The change in PBS to Tris buffer represents a change in FORMULATION. A formulation is defined as:

Pharmaceutical formulation is the multistep process where the active drug is mixed with all other components by considering the factors of particle size, polymorphism, pH, and solubility and becomes the final beneficial medicinal product.

What that means is, the FINAL step in making the drug product going into peoples arms is the mixing of the mRNA loaded LNPs into a stable preparation that is ready to use (or almost ready to use). This is more difficult than it sometimes appears. For ALL parenteral products (including this one), the final product must be:

1. sterile

2. free from pyrogens (like endotoxin)

3. free from visible particulate matter

4. stable (both chemically AND physically including effects of pH, temperature, light, shear, metallic ions, oxygen, etc)

5. isotonic (having the same “tone” or osmotic pressure as the body) and similar pH to the body

6. compatible with diluents if further dilution is required (as was the case with the original purple tops vials)

As we will see Pfizer/BioNTech had considerable issues regarding the particulate matter and stability in their PBS formulation.

The EMA describes the final formulation steps

So after filtration and buffering (to remove the ethanol that the LNPs were stored in), the RNA is measured in the LNPs, then the LNPs are concentrated and buffered in PBS. Sucrose is then added.

Why sucrose? It keeps the LNPs whole during the deep freezing process, because it reduces the ice crystal formation compared to water, and improves the freeze-thaw stability of the LNPs. Then it is filtered into the vials, a visual inspection is done, the vials are labelling and flash frozen at those -60-80C. Then packaging and shipping.

Pfizer tells the regulators just before they are authorized that OOOPs, there are particulates in the vials

Oh but don’t worry, they are just bits of lipids and are of no concern. Pfizer also deflects any toxicity concerns saying that the LNPs were tested for toxicity in 2 studies and the regulators shouldn’t worry. However, it is NOT CERTAIN that the “toxicity” of these particulates were tested since the 2 studies in question used Process 1 lots. And Process 1 lots were never frozen. Only Process 2 lots were frozen and upon thawing is when they found these particulates. So the formulation looking for LNP toxicity may not have had any particulates. Particulates are different than LNP toxicity per se. Please read my substack on Particulates in Pfizer for more. This is how Pfizer/BioNTech distracted from the real issue. And the senior level managers at the EMA/FDA/HC agreed. WHY? This normally would have held back the authorization. In the BC (before Covid) times at least.

THIS IS PFIZER/BIONTECH GAMING THE REGULATORS ROUND 1 ON THE FORMULATION

OK now to Tris. Tris is an ORGANIC buffer while PBS has phosphate and other divalent ions. The formulation chemists at Pfizer finally figured out what Moderna already knew. Tris provided a more stable formulation and was an aldehyde sink. This is further explained in my substack called Lipid Adducts

A Patent by BioNTech using Tris

Our friend Patent Sun on X directed Kevin McKernan and others to this patentt which was filed just at the end of December 2023. Here we get to see what the changes were to the the LNPs using Tris vs PBS.

This image was very striking to me

First of all, the wavy jumpy lines before the main peak represent the truncated and fragmented mRNA, the main peak = intact mRNA and the peak AFTER that are what BioNTech calls “stable folded RNA.” I believe these represents what Moderna calls later eluting peaks which are the lipid adducts.

HOLY TOLEDO!! They were getting stable folded RNA with the PBS buffer as high as 18-21%?? This means upto a fifth of the mRNA was NOT AVAILABLE for translation since these “stable folded RNA” are covalently bound to the lipid and are essentially unusable. A 20% decrease in “dose” so to speak.

Using Tris instead of PBS buffer increases stability and prevents increases in particle size after freezing

Furthermore, using PBS (as described below) causes an INCREASE in PARTICLE size when it is frozen and then thawed. This is what is causing those visible particulates that Pfizer disclosed just before their product was about to be authorized.

Oh and here is evidence that upto a 30% increase in diameter of the LNPs after freezing will be tolerated using the Tris buffer, though more preferably 20% or less. This means the sizes of the LNPs after a freeze-thaw cycle must have increased much more than 30%. They must have been 500nm or much greater given what I read in other parts of the patent. That is a big nanoparticle. If there were larger ones like 1500 nm or aggregates, maybe they could block off capillaries? Just a thought.

What exactly is this “stable folded RNA?” I could not find a definition in the patent but if it is folded and stable I don’t think it is available for translation. Like the lipid adducts. Are they toxic? How does the body get rid of this stable folded RNA? Given the amount of DNA in the Pfizer/BioNTech vaccine do we also have “stable folded DNA” as it is likely the same chemical processes are occurring to the DNA. What does this mean for adverse effects?

So overall, changing to Tris was something that improved the vaccine. It improved stability and decreased the risk of particulates which improved one aspect of safety. Now it doesn’t need to be stored at ultra-cold temperatures, it can be frozen at the same temperature as Moderna, and is more stable out of the fridge as well. Furthermore if the stable folded mRNA are equivalent to lipid adducts then the formation of aldehydes (a potential mutagenic chemical) is inhibited since Tris acts as an aldehyde sink. So far, so good. BUT changing to Tris may mean more available modRNA, and the improvement in potential particulate toxicity should also be verified. Is it possible that these purple-topped vials with visible particulates are more dangerous for acute reactions than the Tris grey topped vials? That is something that needs more study.

Pfizer introduces the Tris formulation at the same time the children’s vaccine is authorized

As mentioned at the very beginning, the FDA approved the Tris formulation for both the children’s and adult versions of the vaccine and also changed the color coding from purple tops to grey tops, and the children got orange topped vials.

There is only one problem. The actual STUDY done on children did not use the Tris formulation. It used the PBS formulation. NEJM study on children.

Here is David Wiseman talking about this topic in 2021 at the World Council of Health when this happened. I can’t clip videos yet so start at 32:20 or so.

Video from WCH David Wiseman 2021

And that it was specifically stated by Pfizer that the, in the children’s vaccine, but the, all the studies were done using the old formulation, PBS buffer and not the TRIS buffer that they use, that they’ve now using, not just for children, but for adults. And so why is that important? The [00:33:02] claimed reason for doing this is stability. And you can see that the stability requirements to the old vaccine are quite stringent and quite difficult to manage. You need a very low temperature and anyone who knows about anything like this, we know this is difficult to manage this sort of cold chain problem. [00:33:17] And so what could have happened is that in some cases, people may have got actually a suboptimal dose and by improving the stability, you could either increase the amount of micrograms of mRNA that everyone would get, or people who might’ve got a lower dose because of stability problems. Now will get a higher dose. [00:33:35] So you’re effectively increasing the effective dose in different ways to people who are going to be receiving this vaccine, which of course could have efficacy and safety consequences on a sort of more chemistry level.

THIS IS PFIZER/BIONTECH GAMING THE REGULATORS ROUND 2 ON THE FORMULATION

and it is a bit more serious. How did the FDA let them get away with it as David says in the video? We were outraged then…..still are.

So less particulates and maybe a higher dose and a different distribution profile? We have no idea.

As far as we are concerned, the change to Tris, supported by this patent, and how it was introduced into the vaccines is another Process change just about on the same level as Process 1 to Process 2.

WE CALL IT THE CHANGE FROM PBS TO TRIS PROCESS 3

KNOWINGLY, KNOWINGLY, KNOWINGLY

Pray the rosary