By Frost on Thursday, March 11, 2004 - 05:48 am:
Hello, everybody
Recently I have started working on purification of protein with HPLC. I'm about to search for solvents which do not denature the protein. At the moment we're using Acetonitrile/water . As I'm a greenhorn on this field (student), my bibliography on solvents is still rather short. I've tried the standard books of Snyder and Sadek, but I'm stuck in database research. Does anyone know a decent source for solvent properties, including their impact on protein?
By HW Mueller on Thursday, March 11, 2004 - 07:28 am:
Each protein has its own "character" so this is a huge field, but, generally, proteins donīt like to be removed from their aqueous media at physiological pH and ionic strength. Any book on protein handling, especially on protein purification (Protein Purification..., Janson, Ryden, VCH, 1989 is a good example) gives a wealth of information. Katalogs like that of Pierce also may be instructive. There are some protocols, etc., etc., on the internet.
Incidentally, pure Acetonitrile-H2O solutions will most certainly denature almost any protein.
One point from my own experience: Stay shy of TFA.
By Frost on Wednesday, March 17, 2004 - 02:03 am:
Thank you very much, I will take your post into consideration.