Antibody separation by hydrophobic charge induction chromatography.
Hydrophobic charge induction chromatography using 4-mercapto-ethyl-pyridine as the ligand is an effective method for the separation of antibodies from a variety of feedstocks. Antibodies are adsorbed in physiological conditions without preliminary concentration. Desorption occurs when the pH is lowered, thus inducing an ionic charge of the same sign to the ligand and the antibody. Antibody capture conditions are compatible with crude samples in terms of pH, conductivity, binding capacity and expression level. The final purity of the antibody is feedstock dependent, but can reach levels of purity as high as 98%. Examples of antibody separation are given and ligand structure information discussed.
Boschetti
Egisto
E
BioSepra, Process division of Ciphergen Biosystems, 48 Avenue des Genottes, F-95800 Cergy Pontoise, France. egisto.boschetti@biosepra.com
D000906:Antibodies / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000302:isolation & purification; D008024:Ligands; D011725:Pyridines / Q000737:chemistry*; D011786:Quality Control; D012680:Sensitivity and Specificity; D013455:Sulfur / Q000737:chemistry
C470522:4-mercaptoethylpyridine; D000906:Antibodies; D007074:Immunoglobulin G; D008024:Ligands; D011725:Pyridines; D013455:Sulfur; C023666:pyridine