Fast purification process optimization using mixed-mode chromatography sorbents in pre-packed mini-columns

• Published in: Journal of Chromatography A 2008-01, Vol.1177 (2), p.226-233
• Main Authors: Brenac Brochier, Virginie, Schapman, Anthony, Santambien, Patrick, Britsch, Lothar
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Cattle; Chickens; Chromatography, Affinity; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; HEA HyperCel; High-throughput screening; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Lactalbumin - isolation & purification; Lactoglobulins - isolation & purification; MediaScout MiniChrom columns; MEP HyperCel; Mixed-mode sorbents; Molecular Weight; PPA HyperCel; Proteins; Reproducibility of Results; Sensitivity and Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2007.08.086

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 β-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.