Method of purifying polypeptides by simulated moving bed chromatography

A method of separating an antibody or antibody fragment from at least one immaterial component in a fluid mixture, comprising:

• (a) providing a simulated moving bed (SMB) chromatography apparatus that comprises a plurality of modules in fluid conducting communication with said apparatus, wherein said modules comprise at least one solid phase comprising a ligand for which the antibody or antibody fragment has selective affinity, and wherein said apparatus comprises a plurality of zones through which the modules pass;
• (b) introducing the fluid mixture into a module in an association zone wherein the fluid mixture contacts the solid phase in a countercurrent mode and the antibody or antibody fragment associates with the solid phase;
• (c) introducing a wash buffer into the module comprising the associated antibody or antibody fragment in at least one wash zone, wherein the wash buffer contacts the solid phase and substantially removes at least one immaterial component from said module;
• (d) introducing an eluent into the module comprising the associated antibody or antibody fragment from step (c) in an elution zone, wherein the eluent contacts the solid phase and promotes disassociation of the antibody or antibody fragment from the solid phase;
• (e) removing the antibody or antibody fragment that is substantially separated from at least one immaterial component in said fluid mixture;
• (f) introducing an elution wash buffer into the module from step (e) in an elution wash zone;
• (g) introducing a solution comprising a regenerant into the module from step (f) in a regeneration zone;
• and
• (h) introducing an equilibration buffer into the module in an equilibration zone, wherein the equilibration buffer creates an environment in the module that permits binding of the antibody or antibody fragment to the solid phase comprising the ligand.