Tomorrow’s headline: Hospitals Flooded with Patients Suffering Racing Hearts – Source Traced to Bioengineered Bacteria. Can Experts Reverse the Epidemic Before More Succumb?
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Today both KF and JW ask about an online article do-it-yourself biotech. Is it actually feasible to duplicate big pharma’s success in producing bioengineered insulin and thyroxine in your own laboratory?
For the technically minded, check out the article they refer to at http://www.indiebiotech.com/?p=135.
While I don’t like to say anything is impossible (are not all things possible with God?), I would estimate the odds of successful production of either product in a home laboratory as close to nil. The article author’s description, while fairly detailed, pales in comparison to what would actually be involved. Perhaps a team of motivated, imaginative, unrestricted, well-funded PhD-level biochemists could concoct these proteins in their basement after a decade of research, but as for counting on this option for treating diabetes and hypothyroidism at TEOTWAWKI, I would not even consider it.
The specific deficiencies the article addresses (lack of insulin in diabetes and lack of thyroxine in hypothyroidism) are really quite separate problems. The thyroid issue is far easier to solve, by resorting to animal sources (and if there aren’t enough animal or human sources around, we’re in big trouble). The thyroid issue is also much simpler because the hormone is effective when taken orally, whereas insulin is not absorbable through the GI tract. Animal thyroid works quite well in humans and thyroid pills should be reasonably stable for years (or perhaps decades) to come. Also, if for some reason thyroid hormone cannot be taken orally, it could be injected (as was done in the late 1800s). Thyroid transplants are theoretically possible but with oral replacement therapy readily available, it has not been a necessity.
The insulin situation is a much greater challenge. It is true that insulin is produced biosynthetically. Recombinant DNA technology now allows for the efficient production of human insulin (which is not immunogenic, like bovine or porcine insulin). However, replicating this technology would be prohibitively difficult. Harvesting insulin from animal pancreatic tissue would be problematic enough.
And what if the recombinant DNA-containg E. coli inoculated the GI tract of researchers? If the engineered bacteria became established as the dominant flora and spread to other family members, the secreted hormones could cause devastating effects. Bacteria producing unquantified amounts of levothyroxine within the human colon could easily cause a condition resembling thyroid storm, a potentially fatal condition.
Although I value imaginative solutions to medical problems, I would classify basement lab production of bioengineered hormones as a pipe dream. (Thanks, KF and JW, for asking.)
On the other hand, if SemBioSys wants to share their secrets of transgenic insulin-producing safflower plants with me (genetic freeware?), I’ll be glad to spread the word.
Please read on for comments from the original article. And how about some input from the other professionals on this site?
- Human Insulin and the Safflower (funflowerfacts.com)
Featured image: insuline crystals
About a year ago I read some interesting info on the blood sugar lowering effects of the southern weed Kudzu. I believe the root held the source of the hypoglycemic agent. Are you aware of any research being done on a product containing the Kudzu vine ? I have been tempted to try gathering, grating and drying the root and making an infusion to test. It’s not insulin but who knows what it might bring to the table.
[Reply from Doc Cindy: I cannot find studies that quantify or confirm the effect, but I do find warnings about low blood sugar, so it may have a hypoglycemic benefit, most likely for Type II diabetics. I’d be interested in learning the results of any self-experimentation you may perform.]
Once you have a complete protocol and the materials making insulin is actually a pretty simple process. The big things are that there are materials you’d want to collect/prepare and store ahead of time.
0. a serious power generation set up. No way to do this without power.
1. 293-6e cells set up for suspension culture stored in aliquots in liquid nitrogen storage (see below)
2. DNA engineered to express secreted insulin with a tag and transfection reagent or set up for electroporation. OR already transfected aliquots of the cells. You could have someone make this for you for probably under $10-15K. (heck, for 15K I’d do it for you.)
3. antibiotics for selection – depends on the DNA construct – many are quite stable at -20C or even room temp for a long time
4. 1 liquid nitrogen storage dewar (I saw a thing about making liquid nitrogen at home with homemade equipment at the Maker Fair) and device to make liquid nitrogen
5. many bags or preweighed powdered media and -80C storage for it (it will last a while at 4C but -80 would be good for years)
6. 1 device to make ultrapure and them make it sterile water
7. chromatography materials – e.g. FLAG resin or his tag resin – depends on what tag you put on the DNA construct. Second, maybe third media for further clean up. most of this stuff is good for years stored at room temp, longer at 4C.
8. Buffers as per protocol and to clean the columns (don’t for get imadazole, flag peptide, etc for column elution). Again mostly stable at RT for years.
9. 2 Scales – 1) Analytical for sub gram amount and; 2) for higher weights. Weigh boats/paper is also a plus.
10. Sterile filters. stable for years. I’d get boxes.
11. Pipettes and pipettors. If you get glass seralogical pipettes you can autoclave them and reuse them. Pipette tips. I’d get cases once again – doubt they ever go bad.
12. pumps and tubing and chromatography columns (and fuses for the pumps, you can do this with gravity but its a pain in the but) – oh, and if you’re going to use a gel filteration column buy one poured for you, it possible to pour one yourself that is as good but you need years of experience. If you have the money I might go with a small akta FPLC.
13. 4C cold space – deli fridge is good
14. 1 controlled atmosphere incubator and roller bottles (you could do wave bags… different set up) (come to think of it you’d want a CO2 generator and means to control the conc
15. Stir plates and teflon stir bars
16. protein for quanitation (BSA is fine)
17. UV spec for protein quantitation or reagent for homemade bradford assay and a spec to read it on. These are abundant and available cheap used.
18. 1 experienced protein biochemist / cell biologist or the equivalent to creat the protocol, troubleshoot it and then teach you how to do it yourself.
Don’t know if that helps or just makes it seem worse. Someone above wrote that they had a protocol – where’d you post it? I’d like to take a peek… :>)
[Check out http://www.nobelprize.org/nobel_prizes/medicine/laureates/1923/banting-lecture.html – Doc Cindy]
As I have a daughter with Type 1, I too, have pondered how to make insulin from animal products in extreme emergency situations. In researching this and in response to an email I’d sent, an older research biochemist wrote that he had heard of insulin in a powdered form (“shelf” stable) that could be reconstituted (how I don’t know) and injected by the diabetic. He said I should ask my doctor. My daughter’s diabetes doctor and our family GP had no idea to what he was referring. I could find nothing online even referring to a powdered form.
Are you aware of the existence, now or in the past, of a powdered form of insulin? It sounds as if it would be a good solution for emergencies for diabetics…if it exists.
For a short time the inhaled insulin powder Exubera by Pfizer was on the market, but it never really caught on and was discontinued. Such a formulation may be re-introduced in the future. It was not intended to be reconstituted with fluid for injection.
As an Islet transplanted type 1, I know when my immuno-suppressants give out, I will need insulin, or say bye, bye…..
I started way back on pork/beef insulin…..
Need the recipe to make own…..with inhaled insulin, is that further processed….would likely be less infection producing….
Lots to learn and know…
As the husband of a type 1 diabetic, I find it interesting that there is little information on the expedient production of insulin. I have read that in China in WW2 an Englishman there manufactured insulin for a family member and friends who were type 1 diabetics. On-line research I have done, while limited, usually talks how insulin was first synthesized (by a Canadian?) but never delves into how it was done. While I agree that a high-tech recombinant DNA solution will most likely be impracticable, I feel that it should be possible to make it using animal products, IF the information on how to is out there. Please note I am not suggesting this as a way to avoid purchasing a needed health product, but as an emergency plan to implement in the event of an apocalyptic situation.
Excellent question, Gary. I currently have a recipe for making insulin up for comment among our professionals. It is quite complicated and would be a challenge. Keep an eye out for further updates.
I’m in the midst of doing the same thing for my wife, who’s rather dependent on several different medicines.
First, check with your pharmacy to see how often the insurance will cover the prescription. Depending on your insurance and the prescription, it might be every 28 days, or every 21 – it’s a slow way to build up, but it’s better than nothing. Be careful that you don’t run into an annual maximum, though. Also, check to see how much it is to buy out the whole prescription at once. The price goes down as the number of pills go up, particularly with generics I had one prescription where it was cheaper than the co-pays to buy out the whole six months at once, and that didn’t go through the insurance at all.
If it was a drug I’d wanted to stockpile, I probably could have had another prescription written (be honest with your doctor and pharmacist about what you’re doing and why) and then gotten the regular fills through the insurance and used them to keep the buffer high. Call different pharmacies though, as prices can differ, and those add up when you start talking about a six month supply.
Another option is changing prescriptions a bit. My wife takes one pill where she can take up to 200mg a day, but she usually only takes 100. I’m going to start getting the prescription filled every month regardless. If there’s flexibility in your dosing, work with that, just make sure you get pills that can be cut or have the prescription written specifically for the lower dose pills. Again, a friendly doctor helps wonders. Also, if your dose changes, the insurance companies generally has to fill it again, even if it’s sooner than they normally would. Pick up a month’s supply of your normal dose a couple days before you get the new dose. If you’re on a nice fancy designer drug, obviously, there are fewer options. But drug companies often give samples of these pills to doctors. Ask all your doctors if they get any such samples, and if they will save them for you. Meanwhile, lets all hope the small bits of insurance reform (not health care reform) that got through last long enough for the pre-exisiting condition rule to kick in .
Hi Cynthia, thanks for your feedback!
As far as feasibility is concerned, it’s not so hard as you might imagine. I’d be the first to remind people that biological prototypes seldom work the first, or even the second, time. However, this was achieved in the 80s and is considered a textbook example by now; back then, they didn’t have remotely the resources we have today.
Engineering these pathways is non-trivial, but it’s certainly not outside the reach of an individual or even an amateur anymore. That was part of the thrust of the article in the first place, to highlight how much things have changed.
On a more practical level, the issue of delivery is, as you well note, more problematic with insulin than thyroxine. With insulin, you could potentially use protein fusions to make isolation of the finished product more straightforward with simple equipment, but injection is going to be out of the question due to residual bacterial antigens. I suggested nasal delivery as a possibility; there’s some work in this area already but retention time and pH can have a big impact on delivery efficiency. Also, depending on which species is manufacturing it for you, you may encounter irritation or inflammation due to aforementioned antigens.
Infectious potential of the producing strain is likely to be nil, but to be on the safe side I generally prefer to encourage use of Bacillus subtilis rather than E.coli for bioengineering in the community. In either case, the use of largely incompetent “Lab Strains” means that they’re unlikely to survive in a competitive environment for long. Even if the bacteria do colonise the GI tract, the expression system should be designed to only produce Thyroxine/Insulin in response to an inducer that isn’t found naturally in the gut. Unless people started dosing themselves with the inducer of choice in a manner that would meaningfully reach and affect the bacteria in the lower gut, there should be no side effects to carrying the engineered strains.
For long-term practicality, I far rather a safflower-like solution. Growth and harvesting of plant-produced medicines is a really practical way to produce medicines on a community scale sustainably. However, bacteria have the advantage of immediacy; there’s no need to wait for the seeds to sprout or roots to take, your culture will be ready within a day or so. It’s more labour intensive and complicated but far more amenable to emergency scenarios. Ideally, you’d have both on standby in a resilient community.
Thanks again for your input; I’m really enjoying the feedback of all stripes that the essay generated!
All the best,