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Fractionation of human plasma proteins by hydrophobic interaction membrane chromatography

机译:疏水相互作用膜色谱分离人血浆蛋白

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摘要

This paper discusses the fractionation of human plasma proteins HSA and HIgG by hydrophobic interaction membrane chromatography. A type of microporous polyvinylidine fluoride (PVDF) membrane having 0.1 μm pore size was identified as being suitable for carrying out this separation. This membrane bound HIgG at 1.5 M ammonium sulphate concentration, a condition at which HSA did not. Based on this selective binding resulting from the selective pressure induced by the high anti-chaotropic salt concentration, these human plasma proteins were fractionated. The HIgG binding capacity of the PVDF membrane examined in this study was 42.8 mg/ml at a feed concentration of 0.45 mg/ml. Separation of simulated HSA/HIgG mixtures were carried out in the pulse and step input modes and the HSA and HIgG fractions thus obtained were analysed for purity using affinity chromatography and SDS-PAGE. HSA and HIgG purities were typically in excess of 97-98%.
机译:本文讨论了通过疏水相互作用膜色谱分离人血浆蛋白HSA和HIgG的方法。具有0.1μm孔径的微孔聚偏氟乙烯(PVDF)膜的类型被确定为适合于进行该分离。该膜以1.5 M的硫酸铵浓度结合HIgG,而HSA则没有。基于高抗离液盐浓度引起的选择性压力产生的这种选择性结合,将这些人血浆蛋白分级分离。在0.45 mg / ml的进料浓度下,本研究中检测到的PVDF膜的HIgG结合能力为42.8 mg / ml。以脉冲和步进输入模式分离模拟的HSA / HIgG混合物,并使用亲和色谱和SDS-PAGE分析由此获得的HSA和HIgG馏分的纯度。 HSA和HIgG纯度通常超过97-98%。

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