Dear researchers,
Is there any method or device like Amicon 0.5ml ultracentrifuge (Its highest cut-off is 100kDa and not good for my purpose) that I could purify my small amount of volume nanoparticles that are conjugated with high MW polymer (more than 400kDa)? I tried 0.5ml ultracentrifugation of 135k*g for 1 hrs but the pellets are not obvious based on a small initial amount of lipid particles that contain nucleic acid.
You have a mixture - high MW polymer and high MW polymer - nanoparticle conjugate.
is the difference in MW sufficient to allow a GPC type separation?
are differences in properties (solubility, acid-base, etc) sufficient to allow other traditional separation techniques? (Precipitation, extraction, etc).
are your nanoparticles magnetic? Charged? Electrophoresis could work
Mehrnoosh Rafiei , one more option, you can try to change the lipid nanoparticle-polymer ratio during the synthesis/conjugation: reduce the initial amount/concentration of the polymer. Afterward, density gradient centrifugation also may be an option.
Dear all, it is expected that polymer-NPs conjugation have higher spatial dimensions compared to neat polymer chains, so membrane dialysis may exclude the polymer. My Regards
In my opinion you could try two different approaches:
- Try diafiltration with an amicon stirred cell and 500 KDa cutt off discs (https://www.merckmillipore.com/ES/es/product/Ultrafiltration-Discs-500kDa-NMW,MM_NF-PBVK07610?ReferrerURL=https%3A%2F%2Fwww.google.com%2F)
-Increase the volume of the ultracentrifugation so that your pellets will be bigger.
Dear All, Thank you for your responses. The best way is to increase the reaction efficiency to ensure that all unwanted polymer chains are conjugated. Subsequently, consider that the spatial dimension of the conjugated nanoparticles is greater than that of the polymer residue! Based on these two considerations, the purification efficiency can be increased!
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