From the looks of it it is a secreted protein with disulfide bonds that is processed by proteases cleaving it at a particular point and other enzymes adding sugar residues at particular points. You’re gonna need to make it in a eukaryote through the secretory pathway and it probably needs a suite of modifying enzymes to cleave and glycosylate it properly, but I don’t know if those enzymes are widespread in the eukaryotes or animal-specific. You can always make it in animal cells but that’s expensive compared to fermentation tanks. It also must make its way INTO circulation, implying injection, and I haven’t been able to see how much plasma was injected and if that indicates occasional infusions or regular injections given how quickly it degrades.
EDIT: see downstream reply to ChristianKl for updated information after I followed the citations and looked at the actual papers after work. Looks like the protein is easier to make than I expected (and indeed a lot of eukaryotic proteins CAN be made in bacteria if you have the industrial-scale equipment to do the proper post-processing), but extremely expensive at this time.
You basically CANNOT give proteins other than digestive enzymes orally.
And I do indeed suspect they’ll find some downside to this somewhere, likely in either cancer rates or metabolic issues.
I should go after the actual papers later today or something...
You’re gonna need to make it in a eukaryote through the secretory pathway and it probably needs a suite of modifying enzymes to cleave and glycosylate it properly, but I don’t know if those enzymes are widespread in the eukaryotes or animal-specific.
If the yeast doesn’t produce the enzyme on it’s own why not simply add the gene for it? If the enzyme messes up the yeast too much you can always switch from yeast towards some other unicellular organism. It might cost a bit to do the bioengineering but when it comes to commercial usage of the protein I don’t think that’s an issue.
Okay, I actually went and looked at the papers and followed a chain of citations back until I found where they are actually getting the protein.
Looks like my assessment was unnecessarily pessimistic. Those that I could find the source of their proteins who studied this protein in isolation seem to have bought a recombinant product from Peprotech, a biotech company that sells huge numbers of proteins. Their website (http://www.peprotech.com/en-US/Pages/Product/Recombinant_Human_GDF-11/120-11) seems to indicate that it is being produced in E. coli. I suspect they have altered the gene a bit to only produce the part cut by the protease that is active and may be doing some post-processing to get it folded right since they indicate that it comes as a dimer held together by disulfide bonds which have a hard time forming in bacterial cytosol. I suspect there’s a bunch of industrial techniques and optimization going on in there that we in the research labs don’t bother with in favor of doing things faster with more complicated or versatile small scale systems.
EDIT: and now I found this http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0085890 in which just this year someone managed to screw up the redox metabolism of bacteria such that you can more easily express proteins that form disulfide bonds in the cytosol of bacteria without the need for expensive post-processing to get the bonds right. One of a class of recent modded bacterial strains that are more amenable to expressing eukaryotic proteins at the expense of some metabolic upset. Cool stuff.
EDIT 2: Additionally, someone in my lab is in the process of trying to make large amounts of a couple of novel weird fungal proteins in order to study their activity in vitro. She can’t express them in a fungus (the context in which they fold normally) since they will alter the cell division cycle of the fungi and probably kill them, so she’s expressing them in bacteria. Had a very hard time getting them to both fold correctly and stay stable without degradation. Over the course of more than six months she’s had to rebuild the system she is using to express it several times, and the final successful version involves expressing the protein alongside a binding partner that is required to make it not aggregate into mush immediately upon translation, fused to a tag that makes it more soluble, and using slightly nonstandard extraction chemicals to keep it from denaturing. You probably need to apply a similarly tailored approach to many novel proteins when you yank them across contexts, which you can justify more readily if you stand to make money off it.
On the other hand, I also looked at a number of recent papers studying the effects of this growth factor. All the studies used daily intraperitoneal injections (injections through the abdominal wall into the space around the intestines) for about a month, of 0.1 mg per kilogram of body weight. That might just be because it can be difficult to get lots of needles into a mouse blood vessel in a short period of time though, but it still suggests that maintaining levels needs lots of injection. Also, according to Peprotech, their cheapest cost is $3900 per milligram (at their current levels of production, which is limited and for research purposes). Of course, insulin is probably a better model of a protein product that makes it big time, though not a perfect one since that prevents rather severe issues immediately rather than slowing chronic ones.
As for stuff brought up in this post, I think that it is probably should indeed possible to muck around with yeast until you can get it to process secreted proteins how you want—just taking a lot longer.
Well, it’s a growth factor. Even IGF1 and growth hormone can rejuvenate old tissue, but they don’t make one age any more slowly (though they can make one more robust up to the end).
Do you think that growth factors like these can accelerate aging in the end though? Reductions in growth factor signalling are often associated with increases in longevity, especially since many growth factors increase mTOR signalling (which often results in lowered rates of protein autophagy). I’m not sure how GDF11 would impact mTOR signalling though.
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Edit: Or is it really a growth factor? https://en.wikipedia.org/wiki/GDF11 says that it’s a growth differentiation factor.. And that it negatively regulates neurogenesis.. Hmm..
http://www.uniprot.org/uniprot/O95390
From the looks of it it is a secreted protein with disulfide bonds that is processed by proteases cleaving it at a particular point and other enzymes adding sugar residues at particular points. You’re gonna need to make it in a eukaryote through the secretory pathway and it probably needs a suite of modifying enzymes to cleave and glycosylate it properly, but I don’t know if those enzymes are widespread in the eukaryotes or animal-specific. You can always make it in animal cells but that’s expensive compared to fermentation tanks. It also must make its way INTO circulation, implying injection, and I haven’t been able to see how much plasma was injected and if that indicates occasional infusions or regular injections given how quickly it degrades.
EDIT: see downstream reply to ChristianKl for updated information after I followed the citations and looked at the actual papers after work. Looks like the protein is easier to make than I expected (and indeed a lot of eukaryotic proteins CAN be made in bacteria if you have the industrial-scale equipment to do the proper post-processing), but extremely expensive at this time.
You basically CANNOT give proteins other than digestive enzymes orally.
And I do indeed suspect they’ll find some downside to this somewhere, likely in either cancer rates or metabolic issues.
I should go after the actual papers later today or something...
As a researcher said the other day (I can’t locate the link right now), “In the end, if heart disease doesn’t get you, cancer will.”
If the yeast doesn’t produce the enzyme on it’s own why not simply add the gene for it? If the enzyme messes up the yeast too much you can always switch from yeast towards some other unicellular organism. It might cost a bit to do the bioengineering but when it comes to commercial usage of the protein I don’t think that’s an issue.
Okay, I actually went and looked at the papers and followed a chain of citations back until I found where they are actually getting the protein.
Looks like my assessment was unnecessarily pessimistic. Those that I could find the source of their proteins who studied this protein in isolation seem to have bought a recombinant product from Peprotech, a biotech company that sells huge numbers of proteins. Their website (http://www.peprotech.com/en-US/Pages/Product/Recombinant_Human_GDF-11/120-11) seems to indicate that it is being produced in E. coli. I suspect they have altered the gene a bit to only produce the part cut by the protease that is active and may be doing some post-processing to get it folded right since they indicate that it comes as a dimer held together by disulfide bonds which have a hard time forming in bacterial cytosol. I suspect there’s a bunch of industrial techniques and optimization going on in there that we in the research labs don’t bother with in favor of doing things faster with more complicated or versatile small scale systems.
EDIT: and now I found this http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0085890 in which just this year someone managed to screw up the redox metabolism of bacteria such that you can more easily express proteins that form disulfide bonds in the cytosol of bacteria without the need for expensive post-processing to get the bonds right. One of a class of recent modded bacterial strains that are more amenable to expressing eukaryotic proteins at the expense of some metabolic upset. Cool stuff.
EDIT 2: Additionally, someone in my lab is in the process of trying to make large amounts of a couple of novel weird fungal proteins in order to study their activity in vitro. She can’t express them in a fungus (the context in which they fold normally) since they will alter the cell division cycle of the fungi and probably kill them, so she’s expressing them in bacteria. Had a very hard time getting them to both fold correctly and stay stable without degradation. Over the course of more than six months she’s had to rebuild the system she is using to express it several times, and the final successful version involves expressing the protein alongside a binding partner that is required to make it not aggregate into mush immediately upon translation, fused to a tag that makes it more soluble, and using slightly nonstandard extraction chemicals to keep it from denaturing. You probably need to apply a similarly tailored approach to many novel proteins when you yank them across contexts, which you can justify more readily if you stand to make money off it.
On the other hand, I also looked at a number of recent papers studying the effects of this growth factor. All the studies used daily intraperitoneal injections (injections through the abdominal wall into the space around the intestines) for about a month, of 0.1 mg per kilogram of body weight. That might just be because it can be difficult to get lots of needles into a mouse blood vessel in a short period of time though, but it still suggests that maintaining levels needs lots of injection. Also, according to Peprotech, their cheapest cost is $3900 per milligram (at their current levels of production, which is limited and for research purposes). Of course, insulin is probably a better model of a protein product that makes it big time, though not a perfect one since that prevents rather severe issues immediately rather than slowing chronic ones.
As for stuff brought up in this post, I think that it is probably should indeed possible to muck around with yeast until you can get it to process secreted proteins how you want—just taking a lot longer.
Well, it’s a growth factor. Even IGF1 and growth hormone can rejuvenate old tissue, but they don’t make one age any more slowly (though they can make one more robust up to the end).
Do you think that growth factors like these can accelerate aging in the end though? Reductions in growth factor signalling are often associated with increases in longevity, especially since many growth factors increase mTOR signalling (which often results in lowered rates of protein autophagy). I’m not sure how GDF11 would impact mTOR signalling though.
==
Edit: Or is it really a growth factor? https://en.wikipedia.org/wiki/GDF11 says that it’s a growth differentiation factor.. And that it negatively regulates neurogenesis.. Hmm..