Akta pure 25m fast protein liquid chromatography system

Heavy chain and light chain sequences of the complimentary determinant regions (CDRs) of centi-C10 ScFv was cloned into each constant region of human IgG1 heavy chain, and human kappa constant IGKC, in the mammalian expression pTT5 vector (Licensed from National Research Council of Canada, Toronto Canada), respectively. Centi-C10 was generated in Transient Transfection using human embryonic kidney Expi293F^™ ThermoFisher Scientific (San Diego, California), according to manufacture protocol at Centivax Inc. (South San Francisco). The antibody was then purified using HiTrap MabSelect PrismA protein A column (Cytiva, Marlborough, MA,), pre-equilibrated with at least 5 column volume (CV) of 1x PBS, pH 7.9 (ThermoFisher Scientific), in AKTA pure 25M fast protein liquid chromatography (FPLC) system (Cytiva). Flow rate was adjusted accordingly depending on the protein titer to achieve appropriate dynamic binding capacity.