采用DOE试验方法优化重组人干扰素α2a纯化工艺

Optimization of Purification Process for Recombinant Human Interferron-α2a by Design of Experiment

目的采用DOE试验方法对重组人干扰素α2a纯化工艺参数进行优化,并寻找各参数的操作空间,为实际的产品生产提供指导。方法利用DOE中的中心符合表面设计(CCF)模型,设置两个响应值目标蛋白纯度和得率,对重组人干扰素α2a纯化工艺的疏水层析精纯进行优化并找出操作空间。选取两个工艺参数,即洗脱液B的(NH4)2SO4浓度(0.60M·L-1~0.70M·L-1)和洗脱液C的(NH4)2SO4浓度(0.30M·L-1~0.40M·L-1)。结果从模型的线性、正确性、有效性和重复性判断,所建立的模型拟合度高,可较为准确的预测实验结果。其中,洗脱液B的(NH4)2SO4浓度、洗脱液C的(NH4)2SO4浓度以及各自的平方对响应值有显著影响。同时,利用sweet spot找出关键层析步骤的操作空间。

OBJECTIVE Optimize the key parameters of recombinant human interferron-α2a purification process, find the operating space and provide guidance for the Pharmaceutical production. METHODS Finding the operating space of key purification process of recombinant human interferron-α2a by the central composite face-cen- tered design（CCF）. For the hydrophobic interaction ,input two factors the concentration of ammonium sulfate in buffer B（0. 60M·L^-1 -0.70M·L^-1 ）and the concentration of ammonium sulfate in buffer C（0.30M·L^-1 -0. 40M·L^-1 ）, output two responses Purity and Yield. RESULTS From the regression coefficients, the predicted accuracy, the model validity and the reproducibility, we can draw the conclusion that the model is good fit, so it can be accu- rately predict the experimental results. For the hydrophobic interaction, the concentration of ammonium sulfate in buffer B, The concentration of ammonium sulfate in buffer C and their square items are the obvious significance. At the end, we got the operating space of the purification processes.