Fast purification process optimization using mixed-mode chromatography sorbents in pre-packed mini-columns.
Pre-packed MediaScout MiniChrom columns of 2.5, 5 and 10 mL were investigated for screening three mixed-mode chromatography sorbents (HEA, PPA and MEP HyperCel). Packing performance was of good quality and the three sorbents displayed higher capacity than traditional HIC sorbents in physiological-like conditions. Each sorbent offered a unique selectivity. Bovine beta-lactoglobulin was partially purified after loading milk whey directly on HEA HyperCel sorbent. The combination of small pre-packed columns and SELDI-MS appeared to be a valuable strategy for high-throughput screening of chromatography sorbents and for enabling rapid process development and optimization.
Brenac Brochier
Virginie
V
Pall BioSepra, PALL France, 48 Avenue des Genottes, Cergy St. Christophe, France. virginie_brochier@europe.pall.com
Schapman
Anthony
A
Santambien
Patrick
P
Britsch
Lothar
L
D000818:Animals; D002417:Cattle; D002645:Chickens; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007768:Lactalbumin / Q000302:isolation & purification; D007782:Lactoglobulins / Q000302:isolation & purification; D008970:Molecular Weight; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D007782:Lactoglobulins; D007768:Lactalbumin