There is another Covid-19 peptide vaccine developed by a Dr. Winfried Stöcker. He injected it into ≥64 volunteers, and the results he published look promising. They show both a good level of IgA, IgG and IgM antibodies and ≥ 94% neutralization for the vast majority of the test subjects. According to him (last paragraph of his blog post), none of the test subjects have reported any relevant adverse symptoms.
Man nehme dreimal 15 Mikrogramm rekombinante RBD der S1-Untereinheit (Arg319-Phe541) für eine Person. Als Adiuvans habe ich Alhydrogel von InvivoGen verwendet: Ordentlich durchschütteln und davon 200 Mikroliter mit der Tuberkulinspritze aufziehen. In eine größere Spritze 10 Milliliter Kochsalz aufziehen und die 200 Mikroliter dazugeben, mischen. Davon 500 Mikroliter pro Schuss, mit denen man seine Portion Antigen vermischt. Alles hübsch steril.
I’ve attempted a translation and added some of my own understanding in [square brackets]. Though I’m a German native speaker, I have zero domain knowledge in this field, so please correct me if anything is wrong:
Take three times 15 μg [three doses of 15 μg per person, spaced days apart—see result data linked above for timing] recombinant RBD in the S1 subunit (Arg319-Phe541) for one person. As an adjuvant I used Alhydrogel by InvivoGen: shake thoroughly and then draw 200 μl with a tuberculin syringe. Using a larger syringe, draw 10 ml saline solution and add the 200 μl [of adjuvant], then mix. Of this mix, use 500 μl per shot and mix it with the antigen [the 15 μg recombinant RBD]. Do all this in a sterile way.
If (!) these instructions are exhaustive, it might be easier, though possibly more expensive to produce than RaDVaC. Googling Arg319-Phe541 suggests that 100 μg (6 doses) / 1000 μg (66 doses) of this RBD can be bought for 310 € (52 € per dose) / 1130 € (18 € per dose), though there may be cheaper offers. I have no clue if the Arg319-Phe541 RBD I found here (230-01102-1000-RB listed on biocat.com and raybiotech.com) would be of the right kind to use in a vaccine.
If you have the relevant domain knowledge to evaluate how complete these instructions are or what other risks and benefits compared to RaDVaC this vaccine may have, I’d really appreciate a comment. Likewise (and especially) if you know where to buy the kind of Arg319-Phe541 RBD used for this vaccine.
If you speak German or are comfortable with DeepL: the current edition (06/21) of the German magazine “Der Spiegel” has some more background on its development on pages 44 − 46: PDF (available for free due to misprints).
Edit 1: Detailed serological results (in English) were posted by Dr. Stöcker in May 2020 when he tested the vaccine on himself first.
If (!) these instructions are exhaustive, it might be easier, though possibly more expensive to produce than RaDVaC.
You need to work a lot more sterile when injecting then when you spray something in your nose. I’d rather have a few additional steps with less problems if I mess up some of the steps.
Good point! I’ve attempted to expand on this a bit, and list the advantages that each vaccine currently seems to have over the other:
For RaDVaC:
Extensive Documentation, Whitepaper and reasoning about its development available
Manufacturing does not require a sterile environment
Simpler administration
Has a small community, might be easier to exchange questions and results
Regularly updated (possibly double-edged—seems very useful to keep up with any variant capable of immune escape, but may (?) make it more difficult to estimate efficacy across vaccine generations)
Designed to prevent immune escape, may still work when commercial vaccines become less effective (uses 9 − 13 peptides instead of just targeting the spike protein like other vaccines)
Cheaper ingredients, because the peptides required are shorter
For Dr. Stöcker’s Vaccine:
Test results released so far show very good efficacy and safety (for n=64)
Known-good dosing regimen available
Efficacy can be verified using commercial blood antibody tests
Requires only one peptide (which might be orderable as-is, without custom synthesis) and two passive ingredients
One way to achieve sterility might be to use a self-made glovebox (example tutorial). For extra safety, you could also add an intake fan with a HEPA filter to get a positive-pressure sterile environment, which would still be sterile in case of a small enough leak. Or build a positive-pressure hood—more upfront work, but working in it seems easier.
Personally, the good results and self-verification capability (using antibody tests) would make me prefer Dr. Stöcker’s vaccine despite the extra hassle, as long as we knew what kind of Arg319-Phe541 peptide we need for it.
Edit 1: Added / edited the last two points in the RaDVaC list, thanks to feedback by ChristianKl.
Has a small community, might be easier to exchange questions and results
Given that this community exists it’s likely that they somehow privately share results. It would be really interesting to know more about what’s going on in that community.
as long as we knew what kind of Arg319-Phe541 peptide we need for it.
I understand Arg319-Phe541 to mean the subsection of the spike protein that begins with arginine at position 319 and ends with phenylalanin at position 541. At the moment I don’t immediately find the sequence with googling but it’s worth checking whether 319 is indeed arginine and 541 phenylalanin to check whether this interpretation makes sense.
The problem is that this is 222 amino acids longs which is longer then what the peptide sequencing company sell you so you can’t get them the same way you get the peptide you need for the RaDVaC vaccine.
I think RaDVaC has another advantage. It’s designed to be difficult for the virus to mutate to get immune to it. Having to change 9 different targets is harder for the virus then just having to change the spike protein and given that every approved vaccine targets the spike protein there’s a lot of selective pressure on the virus mutating to get immune to it. The targets are also careful chosen to be conserved and thus less likely to mutate.
Thank you, this is helpful—I’ve edited the parent post to include some of your feedback.
About the community: the only other place I’ve found so far is /r/radvac—though pretty dead, it may be useful to find people connected to the community. This page also mentions a Boston RaDVaC group.
I understand Arg319-Phe541 to mean the subsection of the spike protein that begins with arginine at position 319 and ends with phenylalanin at position 541.
This seems right—the RaDVaC white paper has a nicely formatted copy of the S protein on page 40, and it begins with R / arginine at 319 and ends with F / phenylalanine at 541.
I found 230-01102-1000-RB by RayBiotech as a cheaper option. If you buy 100 µg of it at 308 € (includes 19 % VAT) you end up at 52 € per dose / 154 € per person. Or you buy 1000 µg at 1129 €, which is enough for 22 people, it’s 18 € per dose / 52 € per person. Again, I have no clue about what you have to look out for when buying this peptide (e.g. format, purity, formulation, …?) for an injected vaccine, any info on this would be highly appreciated!
Apart from that, the data released on the Stöcker vaccine shows that shots 1, 2 and 3 were on average 10 and 32 days apart. Storing the peptides for this amount of time does not seem to be trivial, see e.g. the storage requirements from RayBiotech for their Arg319-Phe541:
Upon arrival, the protein may be stored for 2 weeks at 4 °C. For long term storage, it is recommended to store at −20 °C or −80 °C in appropriate aliquots. Avoid repeated freeze-thaw cycles.
Maintaining 4 °C sounds doable with a good fridge and a data logging thermometer. −20 °C is more tricky—maybe use a home freezer (*** is specced at ≤ −18 °C) and add a data logger. If it then turns out that it can’t reach −20 °C, it might be possible to fix that by modding its internal thermostat somehow. Or have access to a lab freezer, or shell out the big bucks (four figures) to buy one.
I am curious about the storage requirements for RaDVaC and couldn’t find out anything specific on the webpage or in the whitepaper. Though the latter does mention some steps that were taken to improve peptide stability, it would be nice to have some info on this.
Maintaining 4 °C sounds doable with a good fridge and a data logging thermometer. −20 °C is more tricky—maybe use a home freezer (*** is specced at ≤ −18 °C) and add a data logger. If it then turns out that it can’t reach −20 °C, it might be possible to fix that by modding its internal thermostat somehow. Or have access to a lab freezer, or shell out the big bucks (four figures) to buy one.
As someone who has worked in the labs a long time, I wouldn’t worry about having to hit exactly −20 °C; that basically just means “freezer temperature”. Lab freezers don’t work any differently than home freezers as far as I can tell, although they do have certain safety features that a home freezer wouldn’t. But the temperature can still vary a few degrees up or down, and it shouldn’t affect your storage much. The (very) general rule of thumb is a difference of +/- 10 °C makes chemical reactions (such as peptide degradation) go 2x faster/slower. So even having to store in a fridge temporarily would only be ~4x faster than a freezer, still maybe good enough for one’s purposes.
The big difference comes for −20 °C vs −80 °C, since there you have a 2^6 or 64-fold rate difference. So something that can last for a month at −80 °C might degrade in half a day in a freezer. Hence the complex supply chains needed for such vaccines.
Thanks for posting this, this looks excellent.
It’s my impression that you can indeed just buy the antigen needed—the lowest price for 1mg I found was around 900€. Allowing for 10% waste, this would cover 20 individuals at 45µg each. I looked at the antigen test results first and was worried that the vaccine wouldn’t perform as well in live tests, but the neutralization results Stöcker posted are quite promising and a Nature study suggests that using 319–545 of the RBD is effective in providing immunity to live virus in primates. I don’t expect the remaining amino acids will affect the results in a major way, so this looks quite promising indeed.
I’ve asked a few people with more domain knowledge to comment on this and depending on their judgment (and access to sterile lab space) I might fund vaccines for myself and friends, given that Germany’s vaccine rollout seems to be taking its time.
I’ve asked a few people with more domain knowledge to comment on this and depending on their judgment (and access to sterile lab space) I might fund vaccines for myself and friends, given that Germany’s vaccine rollout seems to be taking its time.
Not legal advise: It’s my understanding that German law allows you to take things you brew up yourself but only doctors or Heilpraktikers are allowed to give brews like this to other people. If you have a friend who’s a doctors or Heilpraktikers who could take the role might be helpful (and that person should have a better idea about the exact conditions under which he can give out brews).
From what preliminary legal advice I’ve received, I’m allowed to hand it out as a research chemical but anything beyond that might get me into trouble. That sadly limits me to offering it to other nerds who I can reasonably expect to use it for research purposes, but I also highly doubt that anyone outside that cluster would even be interested.
There is another Covid-19 peptide vaccine developed by a Dr. Winfried Stöcker. He injected it into ≥64 volunteers, and the results he published look promising. They show both a good level of IgA, IgG and IgM antibodies and ≥ 94% neutralization for the vast majority of the test subjects. According to him (last paragraph of his blog post), none of the test subjects have reported any relevant adverse symptoms.
He describes the manufacturing in his blog (see translation below):
I’ve attempted a translation and added some of my own understanding in [square brackets]. Though I’m a German native speaker, I have zero domain knowledge in this field, so please correct me if anything is wrong:
If (!) these instructions are exhaustive, it might be easier, though possibly more expensive to produce than RaDVaC. Googling Arg319-Phe541 suggests that 100 μg (6 doses) / 1000 μg (66 doses) of this RBD can be bought for 310 € (52 € per dose) / 1130 € (18 € per dose), though there may be cheaper offers. I have no clue if the Arg319-Phe541 RBD I found here (230-01102-1000-RB listed on biocat.com and raybiotech.com) would be of the right kind to use in a vaccine.
If you have the relevant domain knowledge to evaluate how complete these instructions are or what other risks and benefits compared to RaDVaC this vaccine may have, I’d really appreciate a comment. Likewise (and especially) if you know where to buy the kind of Arg319-Phe541 RBD used for this vaccine.
If you speak German or are comfortable with DeepL: the current edition (06/21) of the German magazine “Der Spiegel” has some more background on its development on pages 44 − 46: PDF (available for free due to misprints).
Edit 1: Detailed serological results (in English) were posted by Dr. Stöcker in May 2020 when he tested the vaccine on himself first.
You need to work a lot more sterile when injecting then when you spray something in your nose. I’d rather have a few additional steps with less problems if I mess up some of the steps.
Good point! I’ve attempted to expand on this a bit, and list the advantages that each vaccine currently seems to have over the other:
For RaDVaC:
Extensive Documentation, Whitepaper and reasoning about its development available
Manufacturing does not require a sterile environment
Simpler administration
Has a small community, might be easier to exchange questions and results
Regularly updated (possibly double-edged—seems very useful to keep up with any variant capable of immune escape, but may (?) make it more difficult to estimate efficacy across vaccine generations)
Designed to prevent immune escape, may still work when commercial vaccines become less effective (uses 9 − 13 peptides instead of just targeting the spike protein like other vaccines)
Cheaper ingredients, because the peptides required are shorter
For Dr. Stöcker’s Vaccine:
Test results released so far show very good efficacy and safety (for n=64)
Known-good dosing regimen available
Efficacy can be verified using commercial blood antibody tests
Requires only one peptide (which might be orderable as-is, without custom synthesis) and two passive ingredients
One way to achieve sterility might be to use a self-made glovebox (example tutorial). For extra safety, you could also add an intake fan with a HEPA filter to get a positive-pressure sterile environment, which would still be sterile in case of a small enough leak. Or build a positive-pressure hood—more upfront work, but working in it seems easier.
Personally, the good results and self-verification capability (using antibody tests) would make me prefer Dr. Stöcker’s vaccine despite the extra hassle, as long as we knew what kind of Arg319-Phe541 peptide we need for it.
Edit 1: Added / edited the last two points in the RaDVaC list, thanks to feedback by ChristianKl.
Given that this community exists it’s likely that they somehow privately share results. It would be really interesting to know more about what’s going on in that community.
I understand Arg319-Phe541 to mean the subsection of the spike protein that begins with arginine at position 319 and ends with phenylalanin at position 541. At the moment I don’t immediately find the sequence with googling but it’s worth checking whether 319 is indeed arginine and 541 phenylalanin to check whether this interpretation makes sense.
The problem is that this is 222 amino acids longs which is longer then what the peptide sequencing company sell you so you can’t get them the same way you get the peptide you need for the RaDVaC vaccine.
Active-Bioscience seems to sell 100 µg of SARS-CoV-2 Spike Glycoprotein-S1 RBD (319-541, biotinylated) recombinant Protein for 1.150 € which gives you enough doses for two people.
I think RaDVaC has another advantage. It’s designed to be difficult for the virus to mutate to get immune to it. Having to change 9 different targets is harder for the virus then just having to change the spike protein and given that every approved vaccine targets the spike protein there’s a lot of selective pressure on the virus mutating to get immune to it. The targets are also careful chosen to be conserved and thus less likely to mutate.
Thank you, this is helpful—I’ve edited the parent post to include some of your feedback.
About the community: the only other place I’ve found so far is /r/radvac—though pretty dead, it may be useful to find people connected to the community. This page also mentions a Boston RaDVaC group.
This seems right—the RaDVaC white paper has a nicely formatted copy of the S protein on page 40, and it begins with R / arginine at 319 and ends with F / phenylalanine at 541.
I found 230-01102-1000-RB by RayBiotech as a cheaper option. If you buy 100 µg of it at 308 € (includes 19 % VAT) you end up at 52 € per dose / 154 € per person. Or you buy 1000 µg at 1129 €, which is enough for 22 people, it’s 18 € per dose / 52 € per person. Again, I have no clue about what you have to look out for when buying this peptide (e.g. format, purity, formulation, …?) for an injected vaccine, any info on this would be highly appreciated!
Apart from that, the data released on the Stöcker vaccine shows that shots 1, 2 and 3 were on average 10 and 32 days apart. Storing the peptides for this amount of time does not seem to be trivial, see e.g. the storage requirements from RayBiotech for their Arg319-Phe541:
Maintaining 4 °C sounds doable with a good fridge and a data logging thermometer. −20 °C is more tricky—maybe use a home freezer (*** is specced at ≤ −18 °C) and add a data logger. If it then turns out that it can’t reach −20 °C, it might be possible to fix that by modding its internal thermostat somehow. Or have access to a lab freezer, or shell out the big bucks (four figures) to buy one.
I am curious about the storage requirements for RaDVaC and couldn’t find out anything specific on the webpage or in the whitepaper. Though the latter does mention some steps that were taken to improve peptide stability, it would be nice to have some info on this.
As someone who has worked in the labs a long time, I wouldn’t worry about having to hit exactly −20 °C; that basically just means “freezer temperature”. Lab freezers don’t work any differently than home freezers as far as I can tell, although they do have certain safety features that a home freezer wouldn’t. But the temperature can still vary a few degrees up or down, and it shouldn’t affect your storage much. The (very) general rule of thumb is a difference of +/- 10 °C makes chemical reactions (such as peptide degradation) go 2x faster/slower. So even having to store in a fridge temporarily would only be ~4x faster than a freezer, still maybe good enough for one’s purposes.
The big difference comes for −20 °C vs −80 °C, since there you have a 2^6 or 64-fold rate difference. So something that can last for a month at −80 °C might degrade in half a day in a freezer. Hence the complex supply chains needed for such vaccines.
Lab freezers have no defrost cycle. Home freezers often do, which prevents ice buildup but also means they don’t maintain temperature.
So that’s why those damn things were always so full of ice! Thankyou, I did not know this before.
This thread was excellent, strong upvotes for both of you.
Thanks for posting this, this looks excellent.
It’s my impression that you can indeed just buy the antigen needed—the lowest price for 1mg I found was around 900€. Allowing for 10% waste, this would cover 20 individuals at 45µg each. I looked at the antigen test results first and was worried that the vaccine wouldn’t perform as well in live tests, but the neutralization results Stöcker posted are quite promising and a Nature study suggests that using 319–545 of the RBD is effective in providing immunity to live virus in primates. I don’t expect the remaining amino acids will affect the results in a major way, so this looks quite promising indeed.
I’ve asked a few people with more domain knowledge to comment on this and depending on their judgment (and access to sterile lab space) I might fund vaccines for myself and friends, given that Germany’s vaccine rollout seems to be taking its time.
Not legal advise: It’s my understanding that German law allows you to take things you brew up yourself but only doctors or Heilpraktikers are allowed to give brews like this to other people. If you have a friend who’s a doctors or Heilpraktikers who could take the role might be helpful (and that person should have a better idea about the exact conditions under which he can give out brews).
From what preliminary legal advice I’ve received, I’m allowed to hand it out as a research chemical but anything beyond that might get me into trouble. That sadly limits me to offering it to other nerds who I can reasonably expect to use it for research purposes, but I also highly doubt that anyone outside that cluster would even be interested.