Comparison of protein A affinity sorbents III. Life time study

• Published in: Journal of Chromatography A 2006-01, Vol.1102 (1), p.224-231
• Main Authors: Hahn, Rainer, Shimahara, Kazumichi, Steindl, Franz, Jungbauer, Alois
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Antibody; Biological and medical sciences; Cell Line; Chromatography, Affinity - methods; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Fundamental and applied biological sciences. Psychology; Glycoproteins; Host cell protein; Immunoglobulin; Leakage; Proteins; Staphylococcal Protein A - chemistry; Staphylococcus protein A
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2005.10.083

Protein A affinity chromatography is a popular purification method for immunoglobulins applied at various scales, ranging from micro-tube up to 1000 l column format. Three novel high capacity protein A affinity chromatography media have been subjected to a lifetime study using 50 consecutive purification cycles of a cell culture supernatant (CCS) containing a monoclonal antibody. Chromatographic conditions followed protocols used in industrial antibody processing, including stripping and cleaning-in-place of the resins. For all three media, no significant loss of purification performance (measured by sodium dodecylsulfate polyacrylamide gel electrophoresis and analytical size-exclusion chromatography (SEC)) could be observed over 50 cycles. Eluate samples were analyzed for leaked protein A and host cell protein (HCP) content. MabSelect SuRe, the first protein A affinity medium compatible with alkaline regeneration conditions, exhibited the lowest leakage levels, in the range of 1–3 ppm. For the media MabSelect Xtra and ProSep-vA Ultra, leakage levels were in the range of 30–40 ppm. Host cell protein content of eluates from MabSelect Xtra and SuRe were between 300 and 700 ppm, whereas for ProSep-vA Ultra 3000–4000 ppm was achieved.