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Purification of IgG from Sera of Rabbit and Guinea Pig by Flow-Through Mode Ion-Exchange Chromatography using DEAE Sepharose Fast Flow Column |
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| Authors: | Ying Wang Peiyin Zhang Shujun Liu Yongsheng Zhang Tiesuo Zhao Wenhui Huang Chunyan He Yongli Yu Liying Wang Min Wan |
| Institution: | 1. Department of Molecular Biology, Norman Bethune College of Medicine, Jilin University, Changchun, 130021, China 3. Department of Nephropathy, The Second Hospital, Jilin University, Changchun, 130021, China 2. Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun, 130021, China |
| Abstract: | A one-step purification process using flow-through mode ion-exchange chromatography (Ft-IEC) was evaluated for the purification of Immunoglobulin G (IgG) from rabbit and guinea pig serum. A simple protein precipitation with saturated ammonium sulfate was used to pretreated plasma samples. Chromatographic separation was carried out on DEAE Sepharose Fast Flow (F.F.) column at a flow rate of 1 mL min?1. Ft-IEC using Tris?CHCl buffer at pH 7.0 allowed the recovery of 22% of the loaded IgG with purity of 95% existed in flowthrough and washing effluents but not in elution effluents. To be compared, bind-elute mode IEC using Tris?CHCl buffer at pH 8.5 based on the routine protocol showed that the recovery of 15% of the loaded IgG with purity of 80% existed in elution effluents. These results indicate that the Ft-IEC is an effective method for purifying IgG from sera of rabbit and guinea pig and may also be suitable for other animal sera by adjusting pH at equilibrium buffer for chromatography column. |
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