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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >A new approach for the removal of protein impurities from purified biologicals using combinatiorial solid-phase ligand libraries
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A new approach for the removal of protein impurities from purified biologicals using combinatiorial solid-phase ligand libraries

机译：使用组合固相配体库从纯化的生物中去除蛋白质杂质的新方法

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摘要

The removal of last impurity traces from a purified protein is generally called polishing. It is an important step in downstream processing since protein impurities may generate undesirable side effects when the preparation is intended for research, diagnostic and more importantly therapeutic applications. Polishing is generally achieved by using orthogonal separation methods to previous steps, the most common being gel permeation chromatography. In spite of its polishing effectiveness, this technique suffers from a poor separation capacity and modest productivity as a result of low speed. Other approaches, for instance, based on anion exchange or on hydrophobic chromatography, that may be optimized for a given process cannot be used as generic methods. This document reports for the first time the use of a combinatorial solidphase peptide library as a general method for the removal of impurity traces. Several examples of impurity trace removal are reported; starting material is either a pure protein spiked with serum proteins or with Escherichia coli extracts or current purified proteins still containing a small percentage of impurities. Among polished proteins are recombinant human albumin expressed in Pichia pastoris and human transferrin purified from whole plasma. This new method is used in neutral or even physiological pH and ionic strength conditions, with a remarkable capability to remove impurities. The process is as rapid as current adsorption chromatography procedures usable for the removal of a large number of protein impurities, with each one present in small amounts, such as host cell proteins.
机译：从纯化的蛋白质中去除最后的杂质痕迹通常称为抛光。这是下游加工中的重要步骤，因为当该制剂用于研究，诊断以及更重要的是治疗应用时，蛋白质杂质可能会产生不良的副作用。通常通过对先前步骤使用正交分离方法进行抛光，最常见的是凝胶渗透色谱法。尽管具有抛光效果，但由于速度低，该技术的分离能力差且生产率适中。例如，基于阴离子交换或疏水色谱法的其他方法可能无法针对给定的过程进行优化，因此不能用作一般方法。该文件首次报道了组合固相肽库作为去除杂质痕迹的通用方法的使用。报告了去除痕量杂质的几个例子。起始材料是掺有血清蛋白或大肠杆菌提取物的纯蛋白，或目前仍含有少量杂质的纯化蛋白。在抛光的蛋白质中有在毕赤酵母中表达的重组人白蛋白和从整个血浆中纯化的人转铁蛋白。此新方法用于中性甚至生理pH和离子强度条件下，具有出色的去除杂质的能力。该过程的速度与当前用于去除大量蛋白质杂质的吸附色谱法一样快，每种杂质均少量存在，例如宿主细胞蛋白质。

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