Screening of effective column rinse solvent for Protein-A chromatography.
Mildly acidic arginine solution is highly effective in elution of bound proteins from Protein-A columns. Although Protein-A is specific in antibody capture, it does bind other proteins, which must then be removed before elution by aqueous arginine solution. If they are not removed, a strong elution property of aqueous arginine solutions will elute the contaminating proteins along with antibodies. Here we have examined various salt solutions as a column rinse solvent. We screened various solvents for their effects on binding of purified antibodies to Protein-A, instead of their effectiveness to elute the bound contaminants. Those solvents that result in a slight flow-through of the antibodies during loading should be effective in eluting non-specifically bound proteins that have weaker affinity for Protein-A than antibodies: namely, if a particular solvent reduces antibody binding to Protein-A, it is expected to be effective in reducing binding of contaminants and hence eluting them. Such screening showed a few compounds, including arginine and sodium acetate, as potential column rinse agents. A combination of arginine and sodium acetate was tested for a few crude materials containing antibodies.
Yumioka
Ryosuke
R
Applied Research Department, Amino Science Laboratories, Ajinomoto, Inc., 1-1 Suzuki-cho, Kawasaki 210-8681, Japan.
Tsumoto
Kouhei
K
Arakawa
Tsutomu
T
Ejima
Daisuke
D
D000906:Antibodies / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001120:Arginine / Q000737:chemistry; D002845:Chromatography / Q000379:methods*; D011485:Protein Binding; D019346:Sodium Acetate / Q000494:pharmacology; D012997:Solvents; D013205:Staphylococcal Protein A / Q000737:chemistry*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D012997:Solvents; D013205:Staphylococcal Protein A; D019346:Sodium Acetate; D001120:Arginine