I am looking for a methodology to remove protein capping from silver nanoparticles synthesized through mycogenic route. I already tried a methodology, but because of centrifugation steps I lost nanoparticles and I could not obtain the concentration that I need to conduct my experiments. I was looking for a different methodology to avoid nanoparticles loss, but I could not find it yet. Can anyone help me? Thank you very much!
Can you describe the methodology you used? What did you use for stabilization?
Nanoparticles are synthesized by a fungus which acts as a reducing and stabilizing agent, so the stabilization is confered by proteins from the fungus. I do not know which proteins are there.
The methodology was: I centrifuged the nanoparticles, collected the pellet (kept supernatant) and boiled it with SDS 1%. Then I centrifuged again, collected the pellet and boiled it with Tris-HCl 60mM. As I used only the pellet, I obtained a very low volume, so I ressuspended it in water (less than the volume of supernatant).
So, I investigated the concentration through NTA (nanoparticles tracking analysis) and I obtained, in the non-capped nanoparticles, a concentration lower than in the capped. I need to have capped and non-capped both in the same concentration. But in the capping removal part of the nanoparticles remain in supernatant (I saw it through NTA), so I cannot obtain the same concentration of the capped nanoparticles.
Thank you for your attention!
Raising temperature is a bad idea. It will favor particle agglomeration. Run the procedure at room temperature and see what happens. It is likely that if sds does not provide stability you will need to add something else along with it.
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