Format

Send to

Choose Destination
J Chromatogr A. 2015 Jul 10;1402:46-59. doi: 10.1016/j.chroma.2015.05.007. Epub 2015 May 23.

Adsorption equilibrium and kinetics of monomer-dimer monoclonal antibody mixtures on a cation exchange resin.

Author information

1
Department of Chemical Engineering, University of Virginia, Charlottesville, VA, United States.
2
MedImmune, Gaithersburg, MD, United States.
3
Department of Chemical Engineering, University of Virginia, Charlottesville, VA, United States. Electronic address: gc@virginia.edu.

Abstract

Adsorption equilibrium and kinetics are determined for a monoclonal antibody (mAb) monomer and dimer species, individually and in mixtures, on a macroporous cation exchange resin both under the dilute limit of salt gradient elution chromatography and at high protein loads and low salt based on batch adsorption equilibrium and confocal laser scanning microscopy (CLSM) experiments. In the dilute limit and weak binding conditions, the dimer/monomer selectivity in 10mM phosphate at pH 7 varies between 8.7 and 2.3 decreasing with salt concentration in the range of 170-230mM NaCl. At high protein loads and strong binding conditions (0-60mM NaCl), the selectivity in the same buffer is near unity with no NaCl added, but increases gradually with salt concentration reaching high values between 2 and 15 with 60mM added NaCl. For these conditions, the two-component adsorption kinetics is controlled by pore diffusion and is predicted approximately by a dual shrinking core model using parameters based on single component equilibrium and kinetics measurements.

KEYWORDS:

Ion exchange; Monoclonal antibodies; Monomer/dimer separation; Protein adsorption equilibrium and kinetics

PMID:
26028510
DOI:
10.1016/j.chroma.2015.05.007
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center