We are conjugating a protein having MW of 74 KDa to the liposomes. After conjugation which is the best method to separate the Unconjugated proteins from the liposomes conjugated with the protein. For gel chromatography which Sephadex will be suitable for this separation ?
Nitin Charbe Hi. most liposomes have a MW of > 10e6 Da. Most size exclusion resins would be able to separate them fully from a 75 kDa protein.
Hi, as presumably there will be a great difference between your the MW of your protein and that of your liposomes, you have plenty of options.
In this respect, please find attached a PDF manual on the principles and methods of SEC (Size Exclusion Chromatography) and the distinct chromatography column fillings offered by GE Healthcare that I found useful when I had to deal with a similar question.
In your particular case, you need to use a filling whose fractionation range includes the MW of the protein (75 kDa) and whose exclusion limit is below the MW of your liposomes. Of the Sephadex series, Sephadex G-100 meets these requirements (https://www.fishersci.es/shop/products/ge-healthcare-sephadex-g-100-media-2/p-3676369#?keyword=sephadex+100), but you could also use instead for instance Superdex 200 or Sephacryl S-200 (page 49 of the aforementioned manual).
Hope that it works and best of lucks,
Juan
Dear Juan, Thank you very much for the explanation. Its really helpful .
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