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Optimized Purity of a Monoclonal Antibody Using Mixed-Mode Chromatography Media
"For more information, visit: http://www.bio-rad.com/yt/CHT-mAbs Dr. Carsten Voß, Application Specialist, Process Chromatography, Bio-Rad Laboratories GmbH, reviews the important factors and parameters when screening multiple chromatography media (resins) for the development of efficient, robust, and economical production processes of biotherapeutics such as monoclonal antibodies. Using a bind-and-elute strategy provides enhanced purification power compared to flow-through modes in this discussion of evaluating multiple mixed-mode resins for the purification of mAbs. CHT™ Ceramic Hydroxyapatite Mixed-Mode Media provided the best monomer recovery in the smallest elution volume at a target purity of 99.5%. Advantages of Using Mixed-Mode Chromatography A variety of advantages of mixed-mode interactions have been observed beyond simply finer discrimination between product and impurities. They include: ■ Greater resolution compared to unimodal interactions ■ Improved yield and activity via advantageous use of charge-charge repulsion between ligand and protein ■ Large design space for protein binding ■ Minimal feed manipulation prior to binding ■ Mild operating conditions preserve activity CHT Ceramic Hydroxyapatite is a spherical, macroporous form of hydroxyapatite (Ca10(PO4)6(OH)2), which is formed from the chemical combination of calcium and phosphate salts. Unlike most other chromatography media, hydroxyapatite is both the matrix and the ligand, providing multiple modes of interaction. Proteins interact at the negatively charged phosphate (P) site via cation exchange and at the positively charged calcium (C) site via metal affinity. Multiple interactions between stationary and mobile phases can lead to unique selectivity and facilitate separation of closely related proteins and contaminants. Bio-Rad Process Separations offers a full range of chromatography solutions for biomolecule purifications. Visit us on the Web to learn more: http://www.bio-rad.com/en-us/category/mixed-mode-resins?WT.mc_id=sm-PPD-ww-ngc_vyt_201600404-pgZVWocObDo"