Improved purification of immunoglobulin G from plasma by mixed‐mode chromatography |
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Authors: | Dong‐Sheng Chai Yan Sun Xiao‐Ning Wang Qing‐Hong Shi |
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Institution: | 1. Department of Biochemical Engineering, Key Laboratory of Systems Bioengineering, School of Chemical Engineering and Technology, Tianjin University, , Tianjin, China;2. Collaborative Innovation Centre of Chemical Science and Engineering, , Tianjin, China;3. Vaccines Research Department 3, Beijing Tiantan Biological Products Co. Ltd, , Beijing, China |
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Abstract: | Efficient loading of immunoglobulin G in mixed‐mode chromatography is often a serious bottleneck in the chromatographic purification of immunoglobulin G. In this work, a mixed‐mode ligand, 4‐(1H‐imidazol‐1‐yl) aniline, was coupled to Sepharose Fast Flow to fabricate AN SepFF adsorbents with ligand densities of 15–64 mmol/L, and the chromatographic performances of these adsorbents were thoroughly investigated to identify a feasible approach to improve immunoglobulin G purification. The results indicate that a critical ligand density exists for immunoglobulin G on the AN SepFF adsorbents. Above the critical ligand density, the adsorbents showed superior selectivity to immunoglobulin G at high salt concentrations, and also exhibited much higher dynamic binding capacities. For immunoglobulin G purification, both the yield and binding capacity increased with adsorbent ligand density along with a decrease in purity. It is difficult to improve the binding capacity, purity, and yield of immunoglobulin G simultaneously in AN SepFF chromatography. By using tandem AN SepFF chromatography, a threefold increase in binding capacity as well as high purity and yield of immunoglobulin G were achieved. Therefore, the tandem chromatography demonstrates that AN SepFF adsorbent is a practical and feasible alternative to MEP HyperCel adsorbents for immunoglobulin G purification. |
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Keywords: | Immunoglobulin G Ligand density Mixed‐mode adsorbent Plasma purification Tandem chromatography |
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