Method of purifying polypeptides by simulated moving bed chromatography
Inventors
Thommes, Jorg • Sonnenfeld, Alan M. • Pieracci, John P. • Conley, Lynn • Bisschops, Marc • Pennings, Maarten
Assignees
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Publication Number
US-8608960-B2
Publication Date
2013-12-17
Expiration Date
Abstract
Provided are methods of separating an immunoreactive compound from at least one immaterial component, using a simulated moving bed (“SMB”) system and a SMB apparatus for use in these methods. Also provided are purified immunoreactive compounds prepared using the SMB methods and apparatus and methods of treatment with the purified immunoreactive compounds.
Core Innovation
The invention relates to a simulated moving bed system for purifying an antibody or an antibody fragment from at least one immaterial component using a plurality of modules with solid phases arranged into zones that the modules pass through. The zones include an association zone, a first wash zone, an elution zone, an elution wash zone, a regeneration zone, and an equilibration zone, and the system is configured so that each of the plurality of modules serves as one of the respective zones.
In the association zone, the solid phase of a first module is in contact countercurrently with a fluid mixture comprising the antibody or antibody fragment and the at least one immaterial component, where the antibody or antibody fragment is bound to the solid phase of the first module. In the first wash zone, a second module’s solid phase is in contact with a wash zone buffer such that the at least one immaterial component is unbound from the solid phase, and in the elution zone, a third module’s solid phase is in contact with an eluent in which the antibody or antibody fragment is included and unbound from the solid phase.
The first wash zone is intermediate between the association zone and the elution zone. An elution wash zone uses an elution wash buffer intermediate between the elution zone and the association zone, a regeneration zone uses a regenerant solution comprising a regenerant intermediate between the elution wash zone and the association zone, and an equilibration zone uses an equilibration buffer intermediate between the regeneration zone and the association zone. The wash zone buffer is different from the regenerant solution and the equilibration buffer, and binding and unbinding is controlled by the sequence of contacting fluids.
Claims Coverage
The independent claim recites six zone-defined modules in a simulated moving bed system and describes countercurrent association with binding followed by wash, elution, elution wash, regeneration, and equilibration using different fluids. The dependent claims further specify the nature of the solid phase and add structural and flow constraints for the zones and system configuration.
Countercurrent simulated moving bed zoned solid-phase contacting for antibody purification
A simulated moving bed system comprising an antibody or antibody fragment; at least one immaterial component; and a plurality of modules each comprising at least one solid phase, wherein the plurality of modules pass through a plurality of zones including an association zone, a first wash zone, an elution zone, an elution wash zone, a regeneration zone, and an equilibration zone, with each module serving as one of the respective zones such that in the association zone the antibody or antibody fragment is bound to the solid phase via countercurrent contact with a fluid mixture, and then is unbound in the elution zone while other zones provide wash, elution wash, regeneration, and equilibration using different buffers or solutions.
Affinity solid phase for binding the antibody or antibody fragment
The simulated moving bed system wherein the solid phase comprises a ligand used for affinity chromatography.
Protein A or Protein G as the solid-phase ligand
The simulated moving bed system wherein the solid phase comprises Protein A or Protein G.
Cation exchange resin as the solid phase
The simulated moving bed system wherein the solid phase comprises a cation exchange resin.
Each zone includes at least one column
The simulated moving bed system wherein each of the plurality of zones comprises at least one column.
Continuous circulation via a fluid conduit connected to modules
The simulated moving bed system further comprising a fluid conduit connected to the plurality of modules, wherein the fluid conduit is configured to continuously circulate a fluid mixture through the system.
Overall, the claim set covers a simulated moving bed antibody purification architecture defined by sequential association, wash, elution, elution wash, regeneration, and equilibration zones with countercurrent binding in the association zone, then recovery by unbinding in the elution zone, together with dependent refinements that define the solid phase, the zone structure, and continuous circulation using a connected fluid conduit.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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