Anybody interested on working on affinity chromatography? I’ve been playing with ion exchange chromatography and have looked longingly at affinity chromatography for a while as it is a bio technique. Now that their is a 200k machine and proprietary media, I’m thinking about jumping. Anybody with me?
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What you got in mind?!?
“stick with me buddy” only lasts till some tech pours a stronger cocktail?
Conceptually pretty simple…
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Really just locking in the polymers for what we want. It makes me sad to see that company charging a fortune, I’ve been thinking about it for a while now because it fits the bill, biochem molecules. It’s not a new idea, it’s been used for a very long time. I’m happy to share my knowledge, but no 1 person will catch up with a large company with plenty of staff.
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It’s been more than twenty years, but I’m not unfamiliar with the technique.
I’ve made and used traditional affinity columns. From purifying antigen through antibody production, purification and binding to agarose. Then chasing my “molecule of choice” in a variety of extracts.
I don’t have the chemistry to “invent” polymers, however as a molecular biologist, I have some understanding of the search space that can be explored using targeted evolution.
Poly-Peptides that selectively bind one cannabinoid vs another can probably be derived computationally at this point using tools or data out of http://compbio.berkeley.edu or one of their collaborators.
I did my first postdoc there.
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Get me in the game coach.
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I have a r&d spot in las vegas specifically for this. I have a PL-GPC- 50 and some proprietary products. Im interested in THIS.
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Whose the true biochemist? CB1 and 2 are GCPRs which have been studied very extensively. Cell-free expression and affinity chromatography are the important components of these studies. Like @cyclopath said above, we would likely just need the polypeptide sequence that selectively binds similarly to the outward face of the receptors. I have access to a pretty decent peptide synthesizer if that would be adequate, but I would like to have a backup plan for cell free expression of CB1 and 2. Anybody got an idea of direction there? Yeast, E. coli, etc? I know CB2 is significantly smaller than CB1.
I’m not a biochemist other than incidental, required, and collaborative. I know chemistry/physics/polymers and have an extremely analytical background.
My buddy is finishing up his ME PhD at UNLV so I love the Vegas spot!
Since I’m not a biochemist, I’m going down a reading wormhole for a few days. Feel free to point me in a direction.
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Phage display: Phage display—A powerful technique for immunotherapy - PMC
might be one way to screen tens of thousands of engineered or evolved polypeptides for the desired binding in an efficient manner.
I believe it would require radiolabled cannabinoids to not alter the binding, but I’ve not done much work with fluorescent labels.
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Very nice reference. I was completely down until they got to prions and I got weirded out for the night. I’m just kidding of course, but I’m going to have to finish reading it in the morning.
I think affinity chromatography will have a different use than crude to isolate. Sure it sounds good, but dollar for dollar does it make sense to do? With affinity, you’re going to need membrane filters to buffer exchange all of your salts out that co elute with your product prior to dry down. You will still need solvent recovery, but on a larger scale now because of how much solvent these systems use. Your solvent isn’t able to be recycled. When I used to do affinity on AAV, loading my column from a 55gal drum took nearly a full day on an Akta pilot. the pilot has multiple wavelength sensors, conductivity, pH, air sensors, bubble traps, and 8 different inlets for solvents so you don’t salt out your lines. Florida don’t give a fuck about water so we were able to bleach our waste and down the drain with it, so we didn’t even have the added cost of chemical hazmat transport of drums of potentially contaminated solvent. It adds a ton of factors that are already solved for in the current scheme that most use a variation of for the mass market.
Where does this make sense to use? I think stripping refined oil of rare minors would be interesting. I also think this would be great for harvesting cannabinoids from something like a yeast or ecoli reactor. But until you’re selling cannabinoids at the umol like other pure and custom chemicals, I don’t think this would financially make sense to do. Maybe this would be good to make reference standards from.
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I’m game.
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I think this is absolutely about cherry picking the cannabinoids of choice.
So it looks like CBD should actually be easiest based on what I’ve seen. It looks like there are already pre-made media and columns for planar diols.
Off to read some more…
I’ve got a wide variety of polymers coming in for benchtop testing on monday. Gonna be fun. Talked to the chemist who invented a few of the polymers and he directed me towards which will be the best for thc remediation.
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