Many protein purification media have large particle sizes or will not mechanically stand excessive pressure without crushing. (Superdex is an example of the latter.) In general, smaller stationary phase particle sizes give rise to higher selectivity and separation efficiency at the expense of flow rate and possibly capacity. Higher pump pressures are required to get flow through columns packed with tiny particles sizes. To borrow an analytical chemistry maxim: speed, resolution, capacity: pick any two.
Roger Rowlett On Apr 20, 2015 6:45 PM, "xaravich ivan" <[email protected]> wrote: > Hi CCP4eans, > > Do you guys have any preference in purifying a protein by SEC in FPLC > system or using a solvent based HPLC system after the initial affinity > column purification. Where would you prefer HPLC purification over standard > FPLC? > I have routinely seen HPLC based purification of organic molecules and > small peptides but not so much of proteins. > What in your experience are the Pros and Cons of each, in the field of > protein purification? > Any suggestions/insights welcome. > > Thanks, > Ivan >
