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Global Journal of Biochemistry. Volume 2, Issue 1 (2011) pp. 1-27
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Review Article
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Free Article
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The mechanism of elution by MgCl2, ethylene glycol and
arginine in affinity chromatography
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Satoshi Ohtakeb, Kouhei Tsumotoc, Masao Tokunagad, Yoshiko Kitae, Tsutomu Arakawaa
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a Alliance Protein Laboratories, Thousand Oaks, CA 91360, USA
b Aridis Pharmaceuticals, 5941 Optical Court San Jose, CA 95138, USA
c Medical Proteomics Laboratory, Institute of Medical Science, The University of Tokyo
4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan
d Applied and Molecular Microbiology, Faculty of Agriculture,
Kagoshima University, Kagoshima 890-0065, Japan
e Department of Pharmacology, KEIO University School of Medicine, Tokyo 160-8582, Japan
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Abstract |
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Affinity chromatography uses specific interactions present between target macromolecules to be purified and ligands that are conjugated to column matrices. These ligands encompass small molecules, such as enzyme inhibitors, substrates or dyes, and macromolecules, including antigens, antibodies, and Protein-A/G/L. The specific molecular interactions are conferred by the complementary interface present between the target molecule and the ligand, which creates a large van der Waals contact between them, and by multiple interaction forces. These forces encompass hydrophobic interaction, electrostatic interaction, hydrogen bonding, and polar interaction, all contributing to high affinity association. Dissociation of the bound target molecule thus often requires a harsh solvent, e.g., extremely low pH, which can lead to conformational changes of the target macromolecule and the macromolecular ligand. Ethylene glycol (EG) and MgCl2 have been used to replace such harsh conditions and to avoid the degradation of the target molecule to be purified. Arginine has recently been added to the list of useful co-solvents for the same purpose. This review describes exemplary applications of these co-solvents in affinity chromatography, interaction modes between the target molecule and the ligand, and the mechanisms of the effects of co-solvents. Such understanding should help expand their applications in affinity chromatography and develop more effective co-solvent systems or newer classes of co-solvents.
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Keywords |
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Co-solvent; Affinity chromatography; Elution; Preferential interaction; Hydrophobic interaction
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