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Separation and Characterization of Monoclonal Antibody and Fragments
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Monoclonal antibodies have been increasingly becoming drug candidates for disease therapeutics. Due to the molecular complexity of monoclonal antibodies, the characterization remains a challenge and required step throughout the development and manufacturing process. In order to determine the efficacy of the molecules, aggregation, heterogeneity such as charge variants, C-terminal lysine processing, deamidation, glycosylation must be screened for their structural and biological changes. Antibody solution kit offers a complete separation solution for monoclonal antibody analysis and characterization. Sepax offers a complete set of tools for monoclonal antibody analysis, including Zenix SEC-300 and Antibodix NP5 columns. Sepax Zenix-300 size exclusion chromatographic (SEC) column is designed for high efficiency and resolution separation of monoclonal antibody monomers, aggregates, fragments such as heavy/light, fab/fc and f(ab')2 fragments. With its uniform, hydrophilic, and neutral nanometer thick films chemically bonded on high purity silica, the non-specific interaction between proteins and column surface is minimized to result in high resolution of monoclonal antibody separation. With volatile mobile phase systems, Zenix-300 is able to separate MAb fragments, which can be analyzed by on-line mass spectrometry. Molecular weight information of MAb fragments is then obtained with SEC-LC/MS work flow. Sepax Antibodix NP5 weak cation exchange column to separate the charge variants. Multiple mobile phase systems were investigated for optimum charge variant separation. With its non-porous polymer bead, the Antibodix WCX is suitable for resolving slightly different structures of monoclonal antibodies within a wide pH range of 2-12. |