I have one nanoformulation prepared in 70% ethanol. In the formulation there are components like free drug, protein bound drug, free protein and undissolved drug exists. I want to separate the protein bound drug (drug entrapped) which is around 100-200 nm in size without any net charge, from other components and have to quantify with the appropriate method. Please suggest the best separation method.
mass spectrometry(MS) is widely used to quantify the amount of protein bound on the surfaces of NP.
Fluroscence spectroscopy (FS)and Circular dichroism( CD)spectroscopy.
CD for the changes seen on the chiral properties of protein and its secondary structure.
FS can be used for the single purified proteins.
Nuclear magnetic resonance spectroscopy for high resolution information about the protein informations.
you can also use mini column centrifugation method for separation of unbound proteins by using appropriate sepadex. And after centrifugation you will get the nanoparticles/microparticles (bounded with proteins)only .
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