Separation of proteins by microcolumn liquid chromatography based on the reversed-phase and size-exclusion principles
Flurer, C.L.; Borra, C.; Andreolini, F.; Novotny, M.
Journal of Chromatography 448(1): 73-86
1988
ISSN/ISBN: 0021-9673 PMID: 3225298 Document Number: 319996
Slurry-packed fused-silica microcolumns of 250 .mu. I.D., are characterized for use in high-performance liquid chromatographic studies of proteins. The present work utilizes the reversed-phase and size-exclusion chromatographic modes for the separation of standard protein mixtures. A 5-.mu.m, 300-.ANG. octyl material is utilized for the reversed-phase studies, and the size-exclusion studies are accomplished with 5-.mu.m diol material of 60-, 100- and 300-.ANG. pore sizes. Column efficiency and packing reproducibility, as well as sample capacity in a micropreparative mode, are discussed. In addition, the inherent mass sensitivity of a microcolumn liquid chromatography system as applied to protein detection is demonstrated.