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首页> 外文期刊>Bioprocess and Biosystems Engineering >Antibiotic purification from fermentation broths by counter-current chromatography: analysis of product purity and yield trade-offs
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Antibiotic purification from fermentation broths by counter-current chromatography: analysis of product purity and yield trade-offs

机译：通过逆流色谱法从发酵液中纯化抗生素：产品纯度和收率的权衡分析

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Counter-current chromatography (CCC) is a low pressure, liquid-liquid chromatographic technique which has proven to be a powerful purification tool for the high-resolution fractionation of a variety of active pharmaceutical compounds. The successful integration of CCC into either existing or new manufacturing processes requires the predictable purification of target compounds from crude, fermentation-derived, feed streams. This work examines the feasibility of CCC for the purification of fermentation-derived erythromycin A (EA) from its structurally and chemically similar analogues. At the laboratory scale, the effect of feed pre-treatment using either clarified, forward extracted (butyl acetate) or back extracted broth on EA separation was investigated. This defined the degree of impurity removal required, i.e. back extracted broth, to ensure a reproducible elution profile of EA during CCC. Optimisation and scale-up of the separation studied the effects of mobile phase flow (2-40 ml·min~(-1)) and solute loading (0.1-10 g) on the attainable EA purity and yield. The results in all cases demonstrated a high attainable EA purity ( > 97% w/w) with throughputs up to 0.33 kg·day~(-1). Secondly, a predictive scale-up model was applied demonstrating, that from knowledge of the solute distribution ratio of EA (K_(EA)) at the laboratory scale, the EA elution time at the pilot scale could be predicted to within 3-10%, depending upon the solute injection volume. In addition, this study has evaluated a "fractionation diagram" approach to visually determine the effects of key operational variables on separation performance. This resulted in accurate fraction cut-point determination for a required degree of product purity and yield. Overall, the results show CCC to be a predictable and scaleable separation technique capable of handling real feed streams.
机译：逆流色谱（CCC）是一种低压液-液色谱技术，已被证明是用于多种活性药物化合物的高分辨率分离的强大纯化工具。要成功地将CCC整合到现有或新的生产工艺中，就需要从发酵产生的粗制原料流中纯化目标化合物。这项工作检查了CCC从其结构和化学相似的类似物中纯化发酵来源的红霉素A（EA）的可行性。在实验室规模上，研究了使用澄清的正萃取（乙酸丁酯）或反萃取肉汤对饲料进行预处理对EA分离的影响。这定义了所需的杂质去除程度，即反萃取的肉汤，以确保在CCC过程中EA的洗脱曲线具有可再现性。分离的优化和放大研究了流动相流量（2-40 ml·min〜（-1））和溶质负载量（0.1-10 g）对可获得的EA纯度和收率的影响。在所有情况下，结果均显示出较高的EA纯度（> 97％w / w），产量高达0.33 kg·day〜（-1）。其次，应用了预测性放大模型，该模型表明，通过了解实验室规模的EA的溶质分布比（K_（EA）），可以预测中试规模的EA洗脱时间在3-10％之内，取决于溶质注入量。此外，这项研究还评估了“分级图”方法，以直观地确定关键操作变量对分离性能的影响。这样就可以准确确定馏分的临界点，以达到所需的产品纯度和收率。总体而言，结果表明CCC是一种能够处理实际进料流的可预测且可扩展的分离技术。

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