Sciencemadness Discussion Board
Not logged in [Login - Register]
Go To Bottom

Printable Version  
Author: Subject: nematocysts venom extraction
thechemlife
Harmless
*




Posts: 6
Registered: 24-9-2011
Member Is Offline


cool.gif posted on 14-12-2011 at 19:50
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.
View user's profile Visit user's homepage View All Posts By User
fledarmus
Hazard to Others
***




Posts: 187
Registered: 23-6-2011
Member Is Offline


[*] posted on 15-12-2011 at 06:25


This might be the preparation you are looking for...

Ramasamy et al, Toxicology Letters (2005), 219


View user's profile View All Posts By User
thechemlife
Harmless
*




Posts: 6
Registered: 24-9-2011
Member Is Offline


[*] posted on 15-12-2011 at 06:51


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?
View user's profile Visit user's homepage View All Posts By User
Paddywhacker
National Hazard
****




Posts: 474
Registered: 28-2-2009
Member Is Offline


[*] posted on 15-12-2011 at 20:56


Here

Attachment: Ramasamy et al., Toxocology Letters (2005), 219.pdf (118kB)
This file has been downloaded 356 times

View user's profile View All Posts By User
thechemlife
Harmless
*




Posts: 6
Registered: 24-9-2011
Member Is Offline


[*] posted on 15-12-2011 at 21:13


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]
View user's profile Visit user's homepage View All Posts By User
fledarmus
Hazard to Others
***




Posts: 187
Registered: 23-6-2011
Member Is Offline


[*] posted on 16-12-2011 at 04:52


...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!
View user's profile View All Posts By User
White Yeti
International Hazard
*****




Posts: 816
Registered: 20-7-2011
Location: Asperger's spectrum
Member Is Offline

Mood: delocalized

[*] posted on 16-12-2011 at 13:09


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
View user's profile View All Posts By User
thechemlife
Harmless
*




Posts: 6
Registered: 24-9-2011
Member Is Offline


[*] posted on 16-12-2011 at 13:57


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 :P I've been using these forums to come up with ideas for a while now, just never had anything interesting to post.
View user's profile Visit user's homepage View All Posts By User
watson.fawkes
International Hazard
*****




Posts: 2793
Registered: 16-8-2008
Member Is Offline


[*] posted on 16-12-2011 at 15:30


Quote: Originally posted by fledarmus  
...and then the material is assayed for activity. Not purity.
I seems that assaying for activity requires a live animal model, presumably a mouse. Is there another way of assaying for activity?
View user's profile View All Posts By User
thechemlife
Harmless
*




Posts: 6
Registered: 24-9-2011
Member Is Offline


[*] posted on 16-12-2011 at 19:09


Normally they use crayfish as the live model not mice.
View user's profile Visit user's homepage View All Posts By User
phlogiston
International Hazard
*****




Posts: 1103
Registered: 26-4-2008
Location: Neon Thorium Erbium Lanthanum Neodymium Sulphur
Member Is Offline

Mood: pyrophoric

[*] posted on 17-12-2011 at 14:25


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.





-----
"If a rocket goes up, who cares where it comes down, that's not my concern said Wernher von Braun" - Tom Lehrer
View user's profile View All Posts By User
VSEPR_VOID
Hazard to Self
**




Posts: 72
Registered: 1-9-2017
Member Is Offline


[*] posted on 13-11-2017 at 09:53


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.



“Keep a goverment poor and weak and it's your servant; when it is rich and powerful it becomes your master.”
―H. Beam Piper
View user's profile View All Posts By User
NEMO-Chemistry
International Hazard
*****




Posts: 865
Registered: 29-5-2016
Location: UK
Member Is Offline


[*] posted on 13-11-2017 at 12:58


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=s&source=web...

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]
View user's profile View All Posts By User

  Go To Top