Chromatography for the Purification of Biomolecules Industrial Case Studies

DISPLACEMENT CHROMATOGRAPHY FOR THE PURIFICATION OF BIOMOLECULES INDUSTRIAL CASE STUDIES 

Purification of an Antigenic Vaccine Protein by Selective Displacement Chromatography 

FIGURE 14 Purification of rHu-BDNF from its variants by displacement chromatography. (Barnthouse et a/.39) (A) Analytical chromatography of feed stock by high temperature reversed-phase acetonitrile gradient. Column 4.6 X 250 mm Vydac C4 sample 50 jj.% injection of rHu-BNDF. (B) Displacement of rHu - BDNF by protamine. Column 4.6 X 500 mm, POROS HS / M loading 20 mg / mL column volume mobile phase 50 mM phosphate buffer, pH 7.0, 500 m/W NaCI displacer 2 mM protamine sulfate. (C) High-temperature reversed-phase assay of three displacement fractions. Overlay of fractions from the early (fraction 16), intermediate (fraction 18), and late (fraction 31) parts of the displacement train. 

FIGURE 15 Preparative displacement chromatography of a 24 mer phosphorothioate oligonucleotide. (Gerstner et a/.,79 with permission from Oxford University Press.) Load 1.2 g (6 mg/mL Cv) column 50 mmD X 100 mmL POROS HQ-M flow rate 150 cm / hr (50 mL/min) mobile phase 100 mM NaCI, 10 mM NaOH, 5% methanol displacer 800 mL of 7 mg/mL dextran sulfate fraction size 14 mL. 

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