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首页> 外文期刊>Biotechnology Progress >3D-liquid chromatography as a complex mixture characterization tool for knowledge-based downstream process development
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3D-liquid chromatography as a complex mixture characterization tool for knowledge-based downstream process development

机译：3D液相色谱法是用于基于知识的下游工艺开发的复杂混合物表征工具

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Knowledge-based development of chromatographic separation processes requires efficient techniques to determine the physicochemical properties of the product and the impurities to be removed. These characterization techniques are usually divided into approaches that determine molecular properties, such as charge, hydrophobicity and size, or molecular interactions with auxiliary materials, commonly in the form of adsorption isotherms. In this study we demonstrate the application of a three-dimensional liquid chromatography approach to a clarified cell homogenate containing a therapeutic enzyme. Each separation dimension determines a molecular property relevant to the chromatographic behavior of each component. Matching of the peaks across the different separation dimensions and against a high-resolution reference chromatogram allows to assign the determined parameters to pseudo-components, allowing to determine the most promising technique for the removal of each impurity. More detailed process design using mechanistic models requires isotherm parameters. For this purpose, the second dimension consists of multiple linear gradient separations on columns in a high-throughput screening compatible format, that allow regression of isotherm parameters with an average standard error of 8%. (c) 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1283-1291, 2016
机译：基于知识的色谱分离工艺开发需要有效的技术来确定产品和要去除的杂质的物理化学性质。这些表征技术通常分为确定分子性质（例如电荷，疏水性和大小）或与辅助材料发生分子相互作用（通常以吸附等温线形式）的方法。在这项研究中，我们证明了三维液相色谱方法在含有治疗性酶的澄清细胞匀浆中的应用。每个分离维数确定与每个组分的色谱行为有关的分子特性。跨越不同分离尺寸的峰与高分辨率参考色谱图的匹配允许将确定的参数分配给伪组分，从而可以确定去除每种杂质的最有希望的技术。使用机械模型进行更详细的过程设计需要等温线参数。为此，第二维包括高通量筛选兼容格式的色谱柱上的多个线性梯度分离，可实现等温线参数回归，平均标准误为8％。 （c）2016美国化学工程师学会生物技术学会。 Prog。，32：1283-1291，2016

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