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首页> 外文期刊>Protein Expression and Purification >Statistical optimization for immobilized metal affinity purification of secreted human erythropoietin from Drosophila S2 cells
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Statistical optimization for immobilized metal affinity purification of secreted human erythropoietin from Drosophila S2 cells

机译：统计优化的固定的金属亲和力从果蝇S2细胞分泌的人类促红细胞生成素的纯化。

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摘要

We used a novel approach to affinity purify human erythropoietin (hEPO) following its secretion from Drosophila melanogaster S2 cells. Immobilized metal affinity purification of hEPO was optimized using a two-step serial statistical optimization strategy. After determining the elution conditions (based on preliminary batch-type purification experiments), the first optimization step considered three purification factors; resin, equilibrium, and washing. The results of this analysis showed that the resin amount was the major factor influencing yield and purity in both model equations and the washing factor lowered the confidence limits of the acquired model equations. The washing conditions were then set based on the results of the first step optimization and the second step then optimized three factors; resin, equilibrium, and elution. The yield and purity of hEPO were then compared following purification using three different approaches; batch-type purification based upon the conditions determined by serial statistical optimization, batch-type purification performed in preliminary experiments, and FPLC column chromatography-type purification. We found that the serial statistical optimization approach provided the best combination of yield and purity. These findings indicate that serial statistical optimization strategies can be successfully employed for immobilized metal affinity protein purification using either batch-type or column approaches. (C) 2002 Elsevier Science (USA). All rights reserved. [References: 23]
机译：我们使用一种新颖的方法亲和纯化人类促红细胞生成素（hEPO）从果蝇S2细胞分泌的促红细胞生成素。使用两步系列统计优化策略优化了hEPO的固定化金属亲和纯化。确定洗脱条件后（基于初步的分批纯化实验），第一个优化步骤考虑了三个纯化因子;树脂，平衡和清洗。分析结果表明，树脂量是两个模型方程式中影响产率和纯度的主要因素，洗涤因子降低了所获得模型方程式的置信度。然后根据第一步优化的结果设置洗涤条件，然后根据第二步优化的三个因素进行设置。树脂，平衡和洗脱。然后使用三种不同的方法纯化后比较hEPO的产量和纯度。根据连续统计优化确定的条件进行分批式纯化，在初步实验中进行分批式纯化以及FPLC柱色谱法纯化。我们发现串行统计优化方法提供了产量和纯度的最佳组合。这些发现表明，串行统计优化策略可成功地用于使用批次类型或色谱柱方法的固定化金属亲和蛋白纯化。 （C）2002 Elsevier Science（美国）。版权所有。 [参考：23]

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