Adsorption of a Humanized Monoclonal Antibody onto Thermoresponsive Copolymer-Grafted Sepharose Fast Flow Sorbents

The batch adsorption behavior of a humanized monoclonal antibody (hIgG2 mAb) with thermoresponsive polymer (TRP)-modified Sepharose Fast Flow sorbents with different compositions of grafted copolymers is described. At high protein loadings, the adsorption with negatively charged copolymer-modified sorbents exhibited S-shaped isotherms in most cases, indicative of unrestricted multilayer adsorption. The adsorption capacity of the negatively charged copolymer-modified sorbents increased with an increase in the applied environmental temperature due to increased protein-sorbent surface hydrophobic and electrostatic interactions. The affinity of the hIgG2 mAb for a positively charged copolymer-grafted sorbent was much lower than that found for the negatively charged copolymer-grafted sorbents at both 20 and 50 °C due to electrostatic repulsive effects. This study has documented that the molecular functionalities of the grafted copolymer can significantly affect the adsorption behavior of this humanized mAb at both 20 and 50 °C with the isothermal dependencies revealing subtle effects due to copolymer composition.