Parameters optimization in chromatography purification of human immunoglobulin for intravenous injection
Li Guanjun, Zhang Xuecheng, Zhang Jiantao, Liu Yafang, Luo Jian
1Research and Development Center, Hualan Biological Engineering Inc., Xinxiang 453003, China; 2State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
Abstract：Objective To optimize application conditions of Fractogel EMD TMAE chromatography medium in purification of human immunoglobulin for intravenous injection (IVIG) using the response surface method, and to establish the best chromatography parameters. Methods Two-factor, 3-level response surface method was used to design the chromatographic parameters pH and conductivity. Biuret method and high performance liquid chromatography were used to determine the protein concentration and the molecular size distribution, respectively. The protein recovery rate and the content of dimer plus monomer were used to evaluate the effect of purification under different parameters. The optimized chromatographic parameters obtained were then verified. Results Optimized conditions of chromatography purification static balance and sample loading were pH5.40 and conductivity 1.05 ms/cm. Under this chromatography condition, using plasma fraction Ⅱ as raw material, protein recovery was (96.60±3.06) %, IgG dimer plus monomer ratio was (99.98±0.02) %, IgA content was (2.60±1.05) mg/L and IgM content was (4.50±1.35) mg/L. Conclusion Fractogel EMD TMAE chromatography optimized by the response surface method can be used for the preparation of IVIG from plasma component Ⅱ.