A mechanistic study of Protein A chromatography resin lifetime

• Published in: Journal of Chromatography A Vol. 1216; no. 31; pp. 5849 - 5855
• Main Authors: Jiang, Canping, Liu, Jing, Rubacha, Michael, Shukla, Abhinav A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 31-07-2009; Amsterdam; New York: Elsevier; Elsevier
• Subjects: Adsorption; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, Affinity - economics; Chromatography, Affinity - instrumentation; Chromatography, Affinity - methods; Column reuse; Column sanitization; Equipment Reuse; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; General pharmacology; Medical sciences; Monoclonal antibodies; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Protein A chromatography; Protein Binding; Proteins; Receptors, Fc - chemistry; Recombinant Fusion Proteins - chemistry; Resin lifetime; Sepharose - analogs & derivatives; Sepharose - chemistry; Sodium Chloride - chemistry; Sodium Hydroxide - chemistry; Staphylococcal Protein A - chemistry; Resin lifetime; Monoclonal antibodies; Column reuse; Column sanitization; Protein A chromatography; Peak resolution; Purification; Adsorption isotherm; Reuse; Monoclonal antibody; Protein A; Langmuir isotherm; Affinity adsorbent; Separation method; Lifetime; Fc-Fragment; Affinity chromatography; Plate efficiency; Isolation; Height equivalent to a theoretical plate; Fusion protein; Breakthrough curve
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2009.06.013

A mechanistic study into Protein A chromatographic resin lifetime limitations is presented. Binding and mass transport properties of two widely used agarose-based Protein A resins were studied to distinguish between the roles of resin fouling due to product/impurity build-up and ligand degradation as contributory factors towards the decline in binding capacity with use. Cycling studies were conducted with and without product loading on the columns to separate out the influence of resin fouling. Ligand degradation under the mildly alkaline conditions used for column regeneration was determined to be the primary cause for Protein A resin capacity decline with usage. The use of lower concentrations of caustic and the use of stabilizing excipients to protect the Protein A ligand during cleaning and sanitization were found to be useful techniques in maintaining column performance. The results presented in this paper provide a clearer understanding of the causative factors that limit Protein A chromatographic resin lifetime. It is anticipated that these findings will assist in the development of more robust and economical downstream manufacturing processes for monoclonal antibody and Fc fusion protein purification.