Polishing of Monoclonal Antibody Using Hydrophobic Interaction Membrane Chromatography

Submitted to AIChE Annual Meeting 2011

Polishing of monoclonal antibody using hydrophobic interaction membrane chromatography

SeungMi Yoo, Raja Ghosh^*

Department of Chemical Engineering, McMaster University, 1280 Main Street West, Hamilton, Ontario L8S 4L7, Canada; Telephone: +1-905-525-9140 ext 27415; Fax: +1-905-521-1350

Protein-A affinity chromatography is the gold standard capture technology used for purification of monoclonal antibodies. The ligand-bound monoclonal antibodies are usually eluted using acidic pH conditions which result in the formation of antibody aggregates. Another problem associated with protein-A affinity chromatography is the leaching of the immunotoxic ligand from the column during antibody elution. Consequently, the monoclonal antibody purified using this technique needs to be polished to remove among other things, leached protein-A and its fragments as well as antibody aggregates. Generally, combination of polishing techniques such as ion-exchange and size exclusion chromatography is used for monoclonal antibody polishing. Recent work carried out in our group [1] and elsewhere [2] has shown that antibody aggregates are more hydrophobic than the monomeric form of the antibody. In a recent paper, it has also been shown that a protein-A-antibody complex is more hydrophobic than an antibody molecule on its own [3]. In the current study, we attempt to combine these effects to develop a hydrophobic interaction membrane chromatography based technique for removing leached ligand and aggregates from protein-A purified monoclonal antibody. The results obtained are discussed.

References:

1. Wang L. Hale G. Ghosh R. Non-size-based membrane chromatographic separation and analysis of monoclonal antibody aggregates, Analytical Chemistry. 28 (2006) 6863-6867

2. Gagnon p. IgG aggregate removal by charged-hydrophobic mixed mode chromatography. Current Pharmaceutical Biotechnology. 4 (2009) 434-9.

3. Sun X. Yu D. Ghosh R. Study of hydrophobic based binding of immunoglobulin G on synthetic membranes. Journal of membrane Science. 344 (2009) 165-171