| 摘要： |
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. |
