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Advance chromatin extraction improves the performance of electropositive mixed-mode chromatography as a capture step and enables its integration with void-exclusion anion exchange chromatography as a two-column-step purification platform for monoclonal antibody production

Liu, Wenshuai, Sun, Yue, Yu, Jianli, Chen, Quan, Bao, Zixian, Fan, Xiying, Liang, Yunlong, Peng, Xinying, Xian, Mo, Nian, Rui
Biochemical engineering journal 2019 v.142 pp. 145-152
DNA, allantoin, anion exchange chromatography, antibody formation, binding capacity, cation exchange chromatography, chromatin, detection limit, fatty acids, histones, immunoglobulin G, monoclonal antibodies, resins, salt concentration
Advance chromatin extraction, based on a fatty acid/allantoin/solid phase system, has been reported to significantly improve the purification performance of protein A chromatography, cation exchange chromatography and electronegative mixed-mode chromatography to capture monoclonal antibodies (mAbs). In this study, the beneficial effects of advance chromatin extraction were evaluated using electropositive mixed-mode chromatography and were demonstrated to significantly enhance the dynamic binding capacity and removal efficiency of various impurities. Moreover, retention of immunoglobulin G (IgG) molecules on electropositive mixed-mode resins uniquely followed two phases, which decreased and then increased along with the increase in salt concentration. The eluted IgG was directly loaded and further polished by void-exclusion anion exchange chromatography without adjusting the sample. After these two chromatographic steps, non-histone host cell protein (nh-HCP) content in the end-product was reduced to < 1 ppm, DNA was < 1 ppb, aggregates were < 0.05%, and histone HCP was below the detection limit, with an overall process yield of 89.6%. A simplified and seamless two-column-step purification platform with electropositive mixed-mode chromatography as the primary capture strategy was established to provide a more efficient and economical alternative to current prevailing protein A affinity chromatography-based processes.