Protein Purification

[Michael Brown]

Hello,

        I've only just become aware of DIYBio, and have recently completed listening to a Podcast; SynBioBeta Podcast #4: DIYBio.  In it, an individual from Cork named Cathal Garvey was trying to make a recombinant protein purification system within the reach of a layperson.  I wish to enquire as to his progress, and would also like to know about the Hackerspace in sydney, as I am an unemployed biotechnologist looking to utilise some lab space as a Hobby.

[Mega [Andreas Stuermer]]

He's active here. I'm sure you'll get response soon ;)

[Cathal Garvey]

Hey Michael! Glad you enjoyed the podcast. :)

I'm still working on it, and if everything goes well and I'm lucky, I
ought to have something soon. Indeed, if it works, my next round of
gene synthesis will be to order a set of enzymes for easy purification,
to sell as kits at a reasonable price, with full "sourcecode" included
and no nonsense like Patents!

Right now, we're stuck with only one or two proteins commonly purified
DIY; thermostable polymerases. That's because they can be prepared by
freezing and boiling cell samples, and using the clear liquid after
filtering or sedimenting cell matter. It's nice that polymerases are
that easy, but we're lagging everywhere else, because purification
systems either suck (expense, difficulty, secret-sauce) or are patented
to the hilt. Hopefully I can change that, soon.

As far as Sydney labspace, I'm not sure! Hopefully someone here will
chip in?

Feel free to call on me for a tour of my very small lab if you're ever
in Cork. ;)

[Mac Cowell]

The CU-Boulder iGEM 2013 team focused on developing diy methods for molecular biology. miniprep column regeneration, gel electrophoresis band extraction, Taq cloning, and three methods for protein purification. One of these latter methods is based on adding a specific repeat motif to your protein of interest that will cause it to precipitate in the presence of calcium. Not sure if the motif effs up the activity of the protein (very possible), but it's an interesting start.

http://2013.igem.org/Team:CU-Boulder/Project/Kit/Purification

Mac

[Cathal Garvey]

Yea, ELPs: under the right salt/temperature conditions, your protein
alone precipitates, and can then usually re-dissolve once purified by a
quick spin cycle. They're great, really trendy, and patented to the
hilt AFAIK.

My design motivations are:
1) Triviality
2) Ubiquity
3) Freedom

So a purification method must be cheap or use ubiquitous tools, resins
or whatever, really easy, and non-patented.

Sadly, almost everything is patented. I hit upon a rare thing that
wasn't patented, and I'm running with it. Of course, some asshat might
sue me anyway because IP or something, but at least I tried; the point
is more to encourage others to work with what I've built and make
better things, help create an ecosystem. You can't do that with a
patented foundation like ELPs. :-/

[Koeng]

wow those actually look really cool. Thanks for sharing the link!

[Nathan McCorkle]

On Tue, Oct 15, 2013 at 11:44 AM, Cathal Garvey
<cathal...@cathalgarvey.me> wrote:
> Yea, ELPs:

What's an ELP? I only recognize this as a music recording term
(extended long play).

[Cathal Garvey (Phone)]

Elastin-like Peptides

--
Sent from my Android device with K-9 Mail. Please excuse my brevity.

[Josiah Zayner]

Here is a question for you Cathal, why would someone use a new system rather than just using reverse phase HPLC or ion exchange column or even a size exclusion column?

I have purchased HPLC pumps on eBay for ~$50 which includes shipping columns can be purchased for <$10. 

You don't even need a pump I have used ion exchange columns with just a syringe. 

Is the Nickel-NTA patent up? It has been more than 20 years since 1987 when the patent was filed (http://brevets-patents.ic.gc.ca/opic-cipo/cpd/eng/patent/1304886/summary.html) and hard to beat it for price, easy of use and reusability.