On Apr 20, 2015 8:28 PM, "Roger Rowlett" <rrowl...@colgate.edu> wrote:
> Depends on what you want to accomplish... If you have a liter of crude > lysate, "capacity" should be one of the choices. A step gradient is fast > but low resolution; a gradient elution has more resolution but will eat > buffer and take much longer. Lower stationary phase particle size to get > more resolution and flow rates must go down, etc etc. Choices must be made > to achieve the desired goal in the desired time with available resources. > > Roger Rowlett > On Apr 20, 2015 8:14 PM, "Christopher Colbert" < > christopher.colb...@ndsu.edu> wrote: > >> Hi Roger, >> >> Which 2 do you pick? >> >> Cheers, >> >> Chris >> >> -- >> Christopher L. Colbert, Ph.D. >> Assistant Professor >> Department of Chemistry and Biochemistry >> North Dakota State University >> P.O. Box 6050 Dept. 2710 >> Fargo, ND 58108-6050 >> PH: (701) 231-7946 >> FAX: (701) 231-8324 >> >> From: Roger Rowlett <rrowl...@colgate.edu> >> Reply-To: Roger Rowlett <rrowl...@colgate.edu> >> Date: Monday, April 20, 2015 7:07 PM >> To: "CCP4BB@JISCMAIL.AC.UK" <CCP4BB@JISCMAIL.AC.UK> >> Subject: Re: [ccp4bb] HPLC vs FPLC for protein purification >> >> 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" <xaravich.i...@gmail.com> 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 >>> >>
