Separation of proteins by reversed-phase high-performance liquid chromatography. I. Optimizing the column.
In the process of developing a new analytical technology (the chromatophoresis process) which couples reversed-phase high-performance liquid chromatography (HPLC) to sodium dodecyl sulfate polyacrylamide gel electrophoresis in a real-time automated system, it was apparent that improvements in resolving power for the first-dimension (HPLC) separation were necessary. The present paper describes the optimization of the column for our initial work on reversed-phase HPLC separations. Polymeric (polystyrene) packings having particle diameters of 5 micron and pore diameters of 300 A were generally superior in terms of resolution, sample recovery and minimization of "ghosting". Optimum column dimensions were 50 x 1.0 mm I.D. for the flow-rates required in our system (10-100 microliter/min).
Burton
W G
WG
Bionovus, Inc., Foster City, CA 94404.
Nugent
K D
KD
Slattery
T K
TK
Summers
B R
BR
Snyder
L R
LR
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D007202:Indicators and Reagents; D008970:Molecular Weight; D010316:Particle Size; D011506:Proteins / Q000302:isolation & purification*; D012822:Silicon Dioxide
D007202:Indicators and Reagents; D011506:Proteins; D012822:Silicon Dioxide