title: Effects of salts on protein-surface interactions: applications for column chromatography.
abstract: Development of protein pharmaceuticals depends on the availability of high quality proteins. Various column chromatographies are used to purify proteins and characterize the purity and properties of the proteins. Most column chromatographies require salts, whether inorganic or organic, for binding, elution or simply better recovery and resolution. The salts modulate affinity of the proteins for particular columns and nonspecific protein-protein or protein-surface interactions, depending on the type and concentration of the salts, in both specific and nonspecific manners. Salts also affect the binding capacity of the column, which determines the size of the column to be used. Binding capacity, whether equilibrium or dynamic (under an approximation of a slow flow rate), depends on the binding constant, protein concentration and the number of the binding site on the column as well as nonspecific binding. This review attempts to summarize the mechanism of the salt effects on binding affinity and capacity for various column chromatographies and on nonspecific protein-protein or protein-surface interactions. Understanding such salt effects should also be useful in preventing nonspecific protein binding to various containers.
authors.affiliation: Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha, Kashiwa 277-8562, Japan. tsumoto@k.u-tokyo.ac.jp
authors.forename: Kouhei
authors.initials: K
authors.lastname: Tsumoto
authors.forename: Daisuke
authors.initials: D
authors.lastname: Ejima
authors.forename: Anna M
authors.initials: AM
authors.lastname: Senczuk
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Kita
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Arakawa
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation; D002852:Chromatography, Ion Exchange / Q000295:instrumentation; D004867:Equipment Design; D057927:Hydrophobic and Hydrophilic Interactions; D008024:Ligands; D008956:Models, Chemical; D009994:Osmolar Concentration; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012492:Salts / Q000737:chemistry*; D013499:Surface Properties; D013678:Technology, Pharmaceutical / Q000295:instrumentation / Q000379:methods*
chemical_list: D008024:Ligands; D011506:Proteins; D012492:Salts