Ultra-performance liquid chromatography/mass spectrometry of intact proteins.
Given that numerous small molecule applications of ultra-performance liquid chromatography (UPLC) have been published, efforts were made to examine the potential of UPLC to enhance the separation of intact proteins. Beginning with typically employed conditions, column temperature and organic solvent were optimized followed by an HPLC vs. UPLC comparison. When applied to a mixture of 10 protein standards, the optimized method yielded improved chromatographic resolution, enhanced sensitivity, and a threefold increase in throughput. Subsequent cell lysate analysis demonstrated no compromise in chromatographic or mass spectral data quality at 1/3 of the original run time.
Everley
Robert A
RA
Commonwealth of Virginia, Division of Consolidated Laboratory Services, 600 N. 5th Street, Richmond, VA 23219, USA.
Croley
Timothy R
TR
D002851:Chromatography, High Pressure Liquid; D019453:Escherichia coli O157 / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D010316:Particle Size; D011506:Proteins / Q000737:chemistry*; D011786:Quality Control; D012015:Reference Standards; D012997:Solvents; D013696:Temperature
D011506:Proteins; D012997:Solvents