thechemlife
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nematocysts venom extraction
I've always had a bit of an obsession with jellyfish and have wanted to investigate them on a more basic level than just looking at the underwater. My
idea is to collect specimen (dead of the beach) remove the tentacles and then extract the venom from them to see if it has any interesting properties
and see how different species venom functions. Problem is I have no idea how to extract the venom from the nematocysts. I've heard that crushing them
in a ball mill releases the toxin which can be collected but am at a lose if this is true and what sort of solvent would be able to isolate the toxin.
Frankly at this point it is just a crazy idea but am looking for some insight on how to do it. If I can't run any decent tests then it'll just be an
interesting sample to have in my collection of weird chemicals and materials.
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fledarmus
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This might be the preparation you are looking for...
Ramasamy et al, Toxicology Letters (2005), 219
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thechemlife
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The article seems perfect but I can't read the part on the extraction. All I can see is the abstract. Does any one have a PDF copy or a way to find
the procedure elsewhere?
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Paddywhacker
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Here
Attachment: Ramasamy et al., Toxocology Letters (2005), 219.pdf (118kB)
This file has been downloaded 908 times
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thechemlife
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Thanks!
ok so from what I gather from the article, the tentacles are kept in seawater to keep them fresh. Then they are removed and loaded into a mini ball
mill of sorts. Basically just a screw on top vial filled with small glass beads. This should rupture the nematocysts and release the venom. When
enough is collected it is put in a centrifuge tub to remove the left over cell tissue from the venom.
[Edited on 16-12-2011 by thechemlife]
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fledarmus
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...and then the material is assayed for activity. Not purity.
I hadn't read far enough down in the article before I posted it to find that their "pure venom" was actually a mixture of at least 12 different
proteins, and that previous researchers had found difficulties in separating the proteins due to the instability of the purified proteins. It has also
been difficult to tease out the biological effects due to the fact that the mixture of proteins could be affecting multiple pathways.
It looks like an interesting area of study - let me know what you find!
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White Yeti
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I think that you would get poor results because jellyfish also have enzymes that are designed destroy the toxins produced after it dies. Complex
cytotoxins are usually biodegraded rapidly (anatoxin A for example). You could give it a shot, but most of the toxins are proteins that denature very
easily, even upon exposure to light.
Welcome to the forum, it's about time you joined!
"Ja, Kalzium, das ist alles!" -Otto Loewi
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thechemlife
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Ya i've been reading a bit more into them and dont think the toxins would make it from collection in florida to extraction in canada. They normally
freeze dry the tentacles until they are needed but i'll have no equipment so it wont really be a possibility. I guess it'll be a while till i can try
this unless I find a way to get jellyfish shipped for cheap.
Also ya I know it took to long  I've been using these forums to come up with
ideas for a while now, just never had anything interesting to post.
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watson.fawkes
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I seems that assaying for activity requires a
live animal model, presumably a mouse. Is there another way of assaying for activity?
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thechemlife
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Normally they use crayfish as the live model not mice.
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phlogiston
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This method is very unlikely to produce any sort of pure venom. It is an extract of the nematocysts, and it will contain thousands of proteins in
varying concentration used by the cells in that tissue. The 12 bands shown in the SDS-Page are just the most abundant ones and it is unknown whether
these are even part of the venom or just some other abundant proteins from that tissue.
BTW for this purpose the 'ball mill' may not be the best method. This method produces quite a bit of heat, and I find that it sometimes inactivates
enzymes I need to extract from tissue. A simpler method that often gives better results (ie more active enzyme) is to freeze the tissue well
(preferably with liquid N2), and then grind it up with an equally cold mortar and pestle. Then, suspend the ground tissue in cold (0 ... 4 deg C)
buffer, allow it some time to dissolve the proteins (perhaps 20 minutes or so) and finally centrifuge to remove cell debris.
An even better method is to use a potter-elvehjem homogeniser, but you probably don't have access to one.
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"If a rocket goes up, who cares where it comes down, that's not my concern said Wernher von Braun" - Tom Lehrer |
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VSEPR_VOID
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Very interesting. I recently finished a book called "venom" and found the whole subject fascinating. it would be interesting to find and milk a cone
snail for its venom. I imagine that it would not be difficult to separate out all the proteins using electrophoresis or chromatography if you started
with the pure venom.
Within cells interlinked
Within cells interlinked
Within cells interlinked
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NEMO-Chemistry
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electrophoresis is the way i would go, i am allergic to bees the Bee venom is supposed to be pretty nasty. From memory its related to Cobra venom.
I know this sounds way off but, public aquariums often breed jelly fish (at least in the UK). I keep tropical marine and freshwater fish, jelly fish
wouldnt be hard to keep as long as you use a round aquarium like a washing machine type config.
If you got lucky at the right time of year, you could raise them from the plankton layer. Decent dissecting microscope to pick them out then grow on.
Or pick a species with longer lasting Venom, i know some are active upto 7 days after they die. Best thing to do is research the nearest places to
you, then find out what species are most common in that area and what time of year.
Keep ex[anding the search area until you hit a common species for an area, that has a decent life span for the venom, or one that is easy enough to
cultivate.
There are plenty marine biology journals and alot of info on jelly fish. Maybe contact a scientist at a public aquarium nearest to you.
they are often friendly and full of decent info, i would mention a desire to cultivate (might want to not mention cutting them up for a bit). The
other source for possible specimens is large marine aquarium shops, choose ones that import living rock (bare), this is literally hewn from the seabed
of various places and shipped for home aquariums. its common to get small jelly fish in the bottom of the poly box at certain times of year. The shops
dispose of them.
https://www.youtube.com/watch?v=Smkt3MDh4n4
note the above is wrong shape aquarium, you need a washing machine drum shape and constant side flow. I will find a vid with one in
This is the shape
https://www.youtube.com/watch?v=V_DPZQegvpM
Actually thats a great channel and if you want to go down the breeding route, its got some good info.
https://www.youtube.com/channel/UCLvqCW0Nci5hz1lopEDzf8g/vid...
I take it your in Canada?
Here you go, these ship anywhere in Canada, they got some great ones.
http://jellyfishaquarium.ca/
I forgot your other question regarding the the sting after death being active etc.
You dont need an animal model, its often done with agar plates and
chemicals can trigger the process, the jelly fish dosnt have to be alive for the nemocysts to fire. A good protocol for testing methods and protocols
incase your stung are here (good pdf)
https://www.google.co.uk/url?sa=t&rct=j&q=&esrc=...
There is also a book on the box jelly fish, its devoted to purifying its toxin
Techniques Applicable for Purifying Chironex Fleckeri (box-jellyfish) Venom
https://books.google.co.uk/books?id=ZtBjnQAACAAJ
So guaranteed there will be papers for it and protocols.
Get access to ref section (if you dont have it) I have put some papers there on extraction of venom. I will post there what i find
This is first one
www.sciencedirect.com/science/article/pii/S1080603200708210
Second in refs
http://www.sciencedirect.com/science/article/pii/S0041010108...
https://doi.org/10.1016/j.toxicon.2004.04.008
Shout if you need more info. Setting up a aquarium etc i can help you alot with if you need it.
[Edited on 13-11-2017 by NEMO-Chemistry]
[Edited on 13-11-2017 by NEMO-Chemistry]
[Edited on 13-11-2017 by NEMO-Chemistry]
[Edited on 13-11-2017 by NEMO-Chemistry]
[Edited on 13-11-2017 by NEMO-Chemistry]
[Edited on 13-11-2017 by NEMO-Chemistry]
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