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I am interested in extracting aminoacids from microalgae after having ruptured its membrane and carried out proteolysis. After some research, I came to the conclusion that ion exchange chromatography is the best for a medium scaled extraction of aminoacids. However, considering that the tool is quite prohibitive, I was wondering if it were possible to make a small scale DIY version of it, in order to, get a feeling of how it works and what parameters it needs. Even though, I am not sure this question can be asked here, I do wonder how I could make a DIY ion-exchange chromatography system. Even though, a straight-forward answer would be appreciated, I would really value just to hear small tips on it.

Other "side" doubts:

  • How can I "stick" the ionic resin to the tube's wall?
  • Do you know of any online sources that will help me give the system the correct pH parameters in order to achieve optimum net charge?
  • Am I not approaching this correctly, is ion exchange chromatography not the solution?

I do apologize if this question isn't meant to be here. If that is the case, I would be grateful to have the question migrated elsewhere. I will only need to extract these 20 aminoacids (which are, basically, the 20 aminoacids in human metabolism).

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  • $\begingroup$ Are you trying to extract free amino acids or proteins? I'm afraid free amino acids would be a very small fraction of the total content of the cells, and you'd have to get rid of a lot of junk before doing ion exchange chromatography. You're definitely going to need some sort of pH meter to ensure the mobile phase is at the correct pH. Salt concentration will also be an issue. The beads which are the stationary phase are just packed into a column, you don't have to adhere them in any way - there's a frit at the bottom that keeps them from falling out, but allows fluid to pass. $\endgroup$ – MattDMo Oct 22 '20 at 18:57
  • $\begingroup$ @MattDMo Free amino acids though I will first add some hydrochloric acid in order to break the proteins. Lipids are also going to be a problem. $\endgroup$ – david david Oct 22 '20 at 19:11
  • $\begingroup$ HCl may help break some proteins up a bit, but it won't digest them down to amino acids alone, if that's what you're trying to do. To prepare your sample, I would first lyse the cells using an appropriate lysis buffer (with protease inhibitors if you wish), then centrifuge the lysate and only analyze the supernatant. That will help get rid of some of the cell walls, DNA, lipids, etc. $\endgroup$ – MattDMo Oct 22 '20 at 19:19
  • $\begingroup$ @MattDMo Yes, I did forget to mention centrifugation. What would you suggest for the chromatography system? ion exchange systems are too expensive ... $\endgroup$ – david david Oct 22 '20 at 19:23
  • $\begingroup$ I've never purchased a system, so I honestly don't have the slightest idea what they even cost. $\endgroup$ – MattDMo Oct 22 '20 at 19:25

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