Preparative and analytical chromatography of pegylated myelopoietin using monolithic media.
Monolithic media were compared with Q- and SP-Sepharose high performance chromatography for preparative purification and with Q- and SP-5PW chromatography for analysis of a pegylated form of myelopoietin (MPO), an engineered hematopoietic growth factor. The use of either monolithic or Sepharose based supports for preparative chromatography produced highly purified pegylated MPO with the monolithic media demonstrating peak resolution and repeatability at flow rates of 1 and 5 ml/min resulting in run times as much as five-fold shorter compared to Sepharose separations. The monolithic disks also resulted in 10-fold shorter run times for the analytical chromatography, however, their chromatographic profiles and peak symmetry were not as sharp compared to their Q-5PW and SP-5PW counterparts.
Pfizer Inc., 700 Chesterfield Parkway West, Chesterfield, MO 63017, USA. troii.hall@pfizer.com
Troii
T
Hall
David C
DC
Wood
Christine E
CE
Smith
D002850:Chromatography, Gel / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D016179:Granulocyte Colony-Stimulating Factor; D016298:Hematopoietic Cell Growth Factors / Q000032:analysis* / Q000302:isolation & purification*; D007377:Interleukin-3; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000302:isolation & purification*; D011994:Recombinant Proteins
D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C404145:myelopoietin; D016179:Granulocyte Colony-Stimulating Factor