I am testing binding affinities of various antibodies on amyloid fibrils & oligomers using surface plasmon resonance (SPR). I am unable to regenerate the surface (ie remove the amyloid) using the usual regeneration reagents (HCl, SDS, etc). Any ideas?
Amyloid and prions are very challenging to remove from CMC surfaces. You may want to use an alternate substrate, such as PEGs. You can also attempt to use amyloid as the immobilized ligand and flow (monovalent?) Ab over the surface. In this situation, regeneration by 0.5-0.05% SDS may work well. Allow for stabilization time post regeneration. Good Luck!
Hi Francis, The best thing to regenerate surfaces is using plasma. It generally burns off all the organic materials attached to it. You will have to keep it in the plasma chamber for atleast 15 mins and then rinse off with water first followed by ethanol. You will have to determine the time in the plasma chamber based on its power.
Hello Francis! Are you trying to regenerate the surface while you are performing the experiment? I mean, do you want to see a curve during the detachment of amyloid from the surface? If this is not the case, and you just want to clean the gold surface of your sensor, maybe ýou could use piranha solution (3:1 mixture of concentrated sulfuric acid with hydrogen peroxide). I used it to regenerate all my sensors. Cover the sensor surface of this solution for a few minutes (about 5 minutes) and then rinse off with water. But be careful, piranha solution is a strong oxidant mixture. And use it only if you need to regenerate completely the gold surface.
Hi. I use piranha solution to clean of the chip that is covered by biomaterials. But, when I put it in solution all of my gold layer is dissolved in solution.
Do you have any idea about it and what I have to do?
Best sincerely
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