Heterogeneity in therapeutic proteins such as monoclonal antibodies can cause immunogenic responses in the patient as well as lower therapeutic efficacy. In order to control the heterogeneous variants of monoclonal antibodies, product characterization and analytical monitoring are required during the manufacturing process and clinical studies. Ion exchange chromatography is commonly used to study the charge heterogeneity of antibodies due to its ease of use, high sensitivity, and compatibility with biological samples. Chromatographic separations of antibodies often suffer from lower resolution due to a higher resistance to mass transfer term which is attributed to their high molecular mass and large stokes radius. Our approach to this problem is through the use of nonporous particles with small diameters to decreases the resistance to mass transfer term, thus improving the resolution. Herein, we present a novel weak anion exchange (WAX) stationary phase synthesized by grafting a WAX monomer onto the surface of 1.5 �m nonporous silica particles by atom transfer radical polymerization. Column performance was evaluated using ovalbumin as well as a therapeutic monoclonal antibody. Better resolution and selectivity were observed in the ovalbumin and monoclonal antibody samples, respectively when compared to a leading commercial ion exchange column.

Silica based tentacle-type weak anion exchange stationary phase for analysis of monoclonal antibody charge heterogeneity

246th National Meeting (2013)

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