Global screening of protein chromatographic behavior on ion exchangers from a complex cell proteome. Towards in silico downstream processing of bioproducts.
Protein separation during ion-exchange chromatography implies complex physicochemical events. This work has evaluated the chromatographic behaviour of a complex cell proteome on commercial agarose-based adsorbents. Various ligand types in the cation- and anion-exchange mode were studied. ANX-Sepharose, a weak anion exchanger, performed similarly to the strong anion exchanger-type materials. Proteomic tools were applied in order to understand protein separation. Experimental evidence showed a correlation between apparent isoelectric point distributions and the mobile phase conductivity. Molecular weight distributions were unaffected by the elution position. On the basis of two-dimensional electrophoresis, operational windows were described having typical minor contaminants. These could be annotated for future implementation of in silico downstream processing.
Cabrera
Rosa
R
Downstream Processing Laboratory, School of Engineering and Science, Jacobs University Bremen gGmbH, Campus Ring 1, D-28759 Bremen, Germany. r.cabrera@iu-bremen.de
Fernandez-Lahore
Marcelo
M
D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008024:Ligands; D008970:Molecular Weight; D011506:Proteins / Q000302:isolation & purification*; D020543:Proteome
D008024:Ligands; D011506:Proteins; D020543:Proteome