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mRNA vaccines have a "raw materials" shortage
Posted by: AllGold
Date: December 11, 2020 07:56PM
Laurie Garrett says on "All in with Chris Hayes" it's probably due to doing so much testing. "We're using up the entire global supply of RNA extraction kits and the key reagents that are necessary to extract nucleic acids from samples in order to tell if you're positive for this virus. You need the same extraction reagents in order to make an mRNA vaccine."



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Re: mRNA vaccines have a "raw materials" shortage
Posted by: NewtonMP2100
Date: December 11, 2020 08:12PM
....can they work on rearranging the RNA.......to create a mutation.....???



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Re: mRNA vaccines have a "raw materials" shortage
Posted by: PeterB
Date: December 11, 2020 08:48PM
Actually I'd thought about that myself too. There are some simple, dirt-cheap ways to safely and inexpensively isolate RNA in a way that would be amenable to testing. My guess is that nobody is using such methods, because there's a vested interest of the companies who manufacture the reagents/kits to continue getting scientists and clinicians to buy their products, which then means that there very well might be a shortage of reagents for other purposes (such as making a vaccine). In other words, it all comes back to money and greed...




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Re: mRNA vaccines have a "raw materials" shortage
Posted by: Ca Bob
Date: December 12, 2020 03:26AM
There are two classical ways of extracting RNA from a mix of other things, but how complicated you have to make it depends on how you synthesize the mRNA that is used in the vaccine. Do they make the mRNA used in the vaccine using standard synthetic chemistry or are they using recombinant DNA technology? Obviously it would be useful if you could just use synthetic techniques because you don't have to worry about removing extraneous proteins.

Curious question.
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Re: mRNA vaccines have a "raw materials" shortage
Posted by: Speedy
Date: December 12, 2020 08:03AM
This was reported at least a month ago. The Trump Administration has been working hard to alleviate this shortage. (My second sentence is a complete fabrication.)



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Re: mRNA vaccines have a "raw materials" shortage
Posted by: pdq
Date: December 12, 2020 08:06AM
A lifetime ago, I used to precipitate DNA out of aqueous solution with a simple squirt of alcohol. Not exactly the same, but I wonder if PeterB or others with more recent experience (sekker, maybe?) know how they do it today (or the various ways they could do it today).
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Re: mRNA vaccines have a "raw materials" shortage
Posted by: PeterB
Date: December 12, 2020 08:50AM
Quote
pdq
A lifetime ago, I used to precipitate DNA out of aqueous solution with a simple squirt of alcohol. Not exactly the same, but I wonder if PeterB or others with more recent experience (sekker, maybe?) know how they do it today (or the various ways they could do it today).

RNA is harder because it is much more susceptible to breakdown from RNAses (enzymes that break down RNA) and is inherently chemically less stable.

I'm curious what Ca Bob considers to be the two "classical ways", I can guess what at least one of the two of them is. What I was thinking of is a super-fast, quick and dirty way to extract, which would be fine for doing something like PCR ... where you don't necessarily need the highest quality material to work with.

My guess is that they are making the mRNA for vaccines the traditional way, that is, recombinant expression-- rather than completely artificial synthesis, because I would think the latter would be both cost-prohibitive and too difficult to scale up.




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Re: mRNA vaccines have a "raw materials" shortage
Posted by: sekker
Date: December 12, 2020 01:06PM
Yes, synthetic mRNA synthesis using recombinant systems is the production method.

Actually, RNA is far more sensitive to the ubiquitous RNA degradation enzymes. But chemically, RNA is actually more stable than DNA. Once made, and stored properly (this is a BIG if), RNA can be functional for a decade or more.

The bottom line is that there can be some unexpected supply chain issues in molecular biology. For example, when the auto industry came to a major pause during the Great Recession, DNA oligo synthesis was ALSO paused. Why? Turns out, one of the byproduct chemicals of all of the plastic auto seat fabrics is critical for DNA oligo synthesis. No cars, no need for this fabric, no chemical byproduct.

Similarly, everything is balanced. We are now asking for 10x, 100x more out of the supply chain. We will get there in a few months, but maybe not by Feb 2021.
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Re: mRNA vaccines have a "raw materials" shortage
Posted by: neophyte
Date: December 12, 2020 02:04PM
Do it the old-fashion way - hot acidic phenol extraction!
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Re: mRNA vaccines have a "raw materials" shortage
Posted by: PeterB
Date: December 12, 2020 03:45PM
Quote
sekker
Yes, synthetic mRNA synthesis using recombinant systems is the production method.

Actually, RNA is far more sensitive to the ubiquitous RNA degradation enzymes. But chemically, RNA is actually more stable than DNA. Once made, and stored properly (this is a BIG if), RNA can be functional for a decade or more.

Really? I was always taught that RNA is chemically less stable than DNA because of the 2' hydroxyl and that, under most conditions, RNA is single-stranded and therefore the nitrogenous bases can be chemically attacked as well. DNA, once purified, is stable for decades, possibly centuries.

[www.ebi.ac.uk]

And as for purification, I presume that one of CA Bob's two "classical ways" is probably Trizol or a variant thereof (for example, neophyte's hot acidic phenol) and perhaps the second being a column or CTAB purification or again some variant thereof. But if all you need is small amounts of RT-PCR-ready RNA, there are even simpler, cheaper, faster ways to do it...




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