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Dextran-Coated Magnetic Supports Modified with a Biomimetic Ligand for IgG Purification

机译:用仿生配体修饰的葡聚糖涂层磁性载体,用于IgG纯化

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Dextran-coated iron oxide magnetic particles modified with ligand 22/8, a protein A mimetic ligand, were prepared and assessed for IgG purification. Dextran was chosen as the agent to modify the surface of magnetic particles by presenting a negligible level of nonspecific adsorption. For the functionalization of the particles with the affinity ligand toward antibodies, three methods have been explored. The optimum coupling method yielded a theoretical maximum capacity for human IgG calculated as 568 ± 33 mg/g and a binding affinity constant of 7.7 X 10~4 M~(-1). Regeneration, recycle and reuse of particles was also highly successful for five cycles with minor loss of capacity. Moreover, this support presented specificity and effectiveness for IgG adsorption and elution at pH 11 directly from crude extracts with a final purity of 95% in the eluted fraction.
机译:制备了用配体22/8(一种蛋白A模拟配体)修饰的葡聚糖包被的氧化铁磁性颗粒,并对其进行了IgG纯化评估。选择葡聚糖作为通过提供可忽略的非特异性吸附水平来修饰磁性颗粒表面的试剂。为了用对抗体具有亲和配体的颗粒功能化,已经探索了三种方法。最佳偶联方法得到的人IgG理论最大容量为568±33 mg / g,结合亲和常数为7.7 X 10〜4 M〜(-1)。颗粒的再生,回收和再利用在五个循环中也非常成功,而容量损失很小。此外,这种支持物对直接从粗提物中直接在pH值为11的IgG吸附和洗脱提供了特异性和有效性,洗脱级分的最终纯度为95%。

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