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The performance of different pre-treatment techniques in the isolation and purification of monoclonal IgG antibody

机译:不同预处理技术在单克隆IgG抗体的分离和纯化中的性能

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摘要

In this study, the effects of different sample preparation techniques on the separation of monoclonal antibody IgG1 were investigated experimentally. Monoclonal IgG1 was obtained from hybridoma cell line TB/C3 transfected with bcl-2 carrier plasmid, which was grown in serum-free medium. Three different pre-treatment techniques prior to Protein G affinity chromatography have been used in order to concentrate and partial purify the monoclonal antibody. The pre-treatments researched in this paper are precipitation of the antibody by ammonium sulfate, dilution of the antibody in the binding buffer of affinity chromatography and ultrafiltration through an Amicon Ultra-15 filter with molecular weight cut-off at 100 kDa. Purification through direct application of the antibody onto the Protein G affinity column without pre-treatments was used as a control method. The results indicate that the ultrafiltration through an Amicon filter was an effective method for both concentration and partial purification of the antibodies in laboratory scale.
机译:在这项研究中,实验研究了不同样品制备技术对单克隆抗体IgG1分离的影响。从bcl-2载体质粒转染的杂交瘤细胞系TB / C3获得单克隆IgG1,该质粒在无血清培养基中生长。为了浓缩和部分纯化单克隆抗体,在蛋白G亲和层析之前已使用了三种不同的预处理技术。本文研究的预处理方法包括用硫酸铵沉淀抗体,在亲和色谱结合缓冲液中稀释抗体以及通过截留分子量为100 kDa的Amicon Ultra-15滤器进行超滤。通过不经预处理将抗体直接应用到Protein G亲和柱上的纯化方法用作对照方法。结果表明,通过Amicon过滤器进行超滤是在实验室规模上浓缩和部分纯化抗体的有效方法。

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