Tryptic digest analysis by comprehensive reversed phasextwo reversed phase liquid chromatography (RP-LCx2RP-LC) at different pH's.
As an alternative to the classical approach of combining strong cation exchange SCX-LC and RP-LC for the separation of complex proteomic samples, this essay describes the online comprehensive RP-LCxRP-LC separation of BSA and human blood serum. High orthogonality and peak capacity are achieved through the application of a significantly different pH in the two dimensions. The coupling of fused-core columns in series ensures high efficiency in the first dimension, while a previously designed interface with parallel second dimension columns further enhances the separation capability of the comprehensive system.
Separation Sciences Group and Pfizer Analytical Research Centre, Ghent University, Ghent, Belgium.
Isabelle
I
François
Deirdre
D
Cabooter
Koen
K
Sandra
Frédéric
F
Lynen
Gert
G
Desmet
Pat
P
Sandra
D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000379:methods; D044967:Serum / Q000737:chemistry / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism*
D010455:Peptides; D020543:Proteome; D012710:Serum Albumin, Bovine; D014357:Trypsin