1. State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190,
China
2. School of Chemical Engineering, University of Chinese Academy of Sciences, Beijing 100049, China
3. China Institute of Veterinary Drug Control, Beijing 100081, China
Abstract:A single step separation protocol was developed for purification of anti-Salmonella pullorum polyclonal immunoglobulins G (IgG) from rabbit serum. According to the difference of isoelectric point (pI) between IgG and the major impurities, the types of ion exchange media were systematically screened by comparison of their purification results. In order to prevent the IgG from denaturation, different stabilizers were screened by differential scanning fluorimetry (DSF). The results showed that pI of the IgG was 6.04?7.08 determined by capillary iso-electric focusing. After cation exchange chromatography with packing materials of CM Sepharose Fast Flow (CM), the purity analyzed by SDS?PAGE was 63.5% and IgG recovery rate measured by high performance size-exclusion chromatography was 15.5%, respectively. In contrast, the purity of IgG was 99.3% and recovery rate was 67.5% after anion exchange chromatography with packing materials of Q Sepharose XL (Q-XL). 200 g/L sorbitol was found to possess the best protection effect. The two thermal denaturation temperatures of IgG were increased by 5.52 and 8.84℃, respectively, and the stability at 70℃ was significantly improved. The results demonstrated that the one step anion exchange chromatography together with 200 g/L sorbitol protection provided a high purity, high recovery rate, and high stability of the IgG. The whole process is facile and efficient.
尤星力 杨延丽 苏志国 张媛 张松平. 鸡白痢沙门氏菌兔抗血清多抗IgG的纯化与稳定[J]. 过程工程学报, 2019, 19(6): 1204-1211.
Xingli YOU Yanli YANG Zhiguo SU Yuan ZHANG Songping ZHANG. Facile purification and stabilization of anti-Salmonella pullorum polyclonal immunoglobulin G. Chin. J. Process Eng., 2019, 19(6): 1204-1211.
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