Elution of antibodies from a Protein-A column by aqueous arginine solutions.
Acidic pH is commonly used to elute antibodies from Protein-A affinity column, although low pH may result in aggregation of the proteins. As an alternative, here arginine was tested as an eluent and compared with a more conventional eluent of citrate. Using purified monoclonal antibodies, recovery of antibodies with 0.1M citrate, pH 3.8, was less than 50% and decreased further as the pH was increased to 4.3. At the same pH, the recovery of antibodies was greatly increased with 0.5M arginine and more so with 2M arginine. Even at pH 5.0, 2M arginine resulted in 31% recovery, although the elution under such condition showed extensive tailing. Such tailing was observed at pH 3.8 when 0.1M citrate was used. Size exclusion analysis indicated that the eluted antibodies were mostly monomeric whether eluted with citrate or arginine. This demonstrates the usefulness of arginine as an efficient eluent for Protein-A chromatography.
Arakawa
Tsutomu
T
Alliance Protein Laboratories, Inc., 3957 Corte Cancion, Thousand Oaks, CA 91360, USA. tarakawa@aol.com
Philo
John S
JS
Tsumoto
Kouhei
K
Yumioka
Ryosuke
R
Ejima
Daisuke
D
D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D001120:Arginine / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D019343:Citric Acid / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012996:Solutions / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D012996:Solutions; D013205:Staphylococcal Protein A; D019343:Citric Acid; D001120:Arginine