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首页> 外文期刊>Separation and Purification Technology >Affinity-enhanced purification of human antibodies by aqueous two-phase extraction
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Affinity-enhanced purification of human antibodies by aqueous two-phase extraction

机译：水相两相萃取亲和增强人抗体的纯化

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摘要

The partition of human antibodies in polyethylene glycol (PEG)/dextran aqueous two-phase systems (ATPS) was investigated using charged, hydrophobic and affinity ligands, either in the form of free ligands or as phase forming components. Free ligands were based on triethylene glycol (TEG) molecules modified with glutaric acid, sulfonate, mercaptoethyl pyridine, aminoquinuclidine and pyrimidine groups. In addition, PEG molecules with a molecular weight of 1000 were modified with amino groups and also used as free ligands. Glutaric acid displayed a high affinity towards the antibodies with a 96% extraction yield and protein purity of 95%. PEG molecules with a molecular weight of 3350 were modified with glutaric acid, amino, mercaptoethyl pyridine, pyrimidine and benzyl groups, and used as phase forming components. Best results were obtained with glutaric acid, amino and benzyl groups with extraction yields higher than 75% and protein purities around 90%. The best performing system was composed of 5% dextran, 8% PEG diglutaric acid, 10 mM phosphate buffer pH 7, which allowed the extraction of 97% of IgG to the upper phase, with a purity of 94% in terms of proteins.
机译：使用带电荷的，疏水的和亲和的配体，以游离配体形式或作为相形成组分，研究了人抗体在聚乙二醇（PEG）/葡聚糖水两相系统（ATPS）中的分配。游离配体基于戊二酸，磺酸盐，巯基乙基吡啶，氨基喹核苷和嘧啶基团修饰的三甘醇（TEG）分子。此外，分子量为1000的PEG分子被氨基修饰，也可用作游离配体。戊二酸显示出对抗体的高亲和力，提取率为96％，蛋白质纯度为95％。用戊二酸，氨基，巯基乙基吡啶，嘧啶和苄基修饰分子量为3350的PEG分子，并用作相形成组分。用戊二酸，氨基和苄基可获得最佳结果，提取率高于75％，蛋白质纯度约为90％。表现最好的系统由5％葡聚糖，8％PEG二戊二酸，10 mM磷酸盐缓冲液pH 7组成，可将97％的IgG提取到上层相中，其蛋白质纯度为94％。

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