MEP chromatography of antibody and Fc-fusion protein using aqueous arginine solution.
MEP HyperCel resin, one of the Protein-A mimetic columns, is designed to bind antibodies at physiological pH and elutes the bound antibodies at mildly acidic pH. We have tested aqueous arginine solution for washing and elution of the resin. To our surprise, bound antibody and Fc-fusion protein eluted at pH 7.0 using 1M arginine solution. Various solvent additives were then examined at pH 7.0. Among the tested additives, urea and arginine were the only additives that were effective in elution. Thus, urea and arginine at low concentrations were effectively used for washing the resin. NaCl and MgCl(2) at 0.1-1M and ethanol at 5-20% were not effective. Based on these observations, it appears that protein binds to MEP resin through both polar and hydrophobic interactions with some contribution of electrostatic interaction, which can be simultaneously reduced by arginine or urea. On the other hand, Mabsorbent, another Protein-A mimetic column, appears to be more non-specific and non-selective.
Arakawa
Tsutomu
T
Alliance Protein Laboratories, Inc., 3957 Corte Cancion, Thousand Oaks, CA 91360-6919, USA. tarakawa2@aol.com
Kita
Yoshiko
Y
Sato
Haruna
H
Ejima
Daisuke
D
D000906:Antibodies / Q000187:drug effects / Q000302:isolation & purification*; D001120:Arginine / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D007141:Immunoglobulin Fc Fragments / Q000187:drug effects / Q000302:isolation & purification*; D019995:Laboratory Chemicals / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000302:isolation & purification*; D012996:Solutions; D014508:Urea / Q000737:chemistry; D014867:Water / Q000737:chemistry
D000906:Antibodies; D007141:Immunoglobulin Fc Fragments; D019995:Laboratory Chemicals; D011993:Recombinant Fusion Proteins; D012996:Solutions; D014867:Water; D014508:Urea; D001120:Arginine