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通过亲和色谱策略纯化天然和重组肽抗体

The Purification of Natural and Recombinant Peptide Antibodies by Affinity Chromatographic Strategies.

作者信息

Ma Hui, O'Kennedy Richard

机构信息

School of Biotechnology and Biomedical Diagnostics Institute, Dublin City University, Glasnevin, Dublin 9, Ireland.

出版信息

Methods Mol Biol. 2015;1348:153-65. doi: 10.1007/978-1-4939-2999-3_15.

Abstract

The purification of peptide antibodies (e.g., IgG, IgY, scFv, and Fab) are described in this chapter. Affinity chromatographic purification, a very convenient and effective antibody purification strategy, is used to isolate peptide antibodies based on specific binding, i.e., binding of the antibody to a column on which its specific ligand is immobilized with subsequent elution of the purified antibody. In addition, the application of purification methods based on the use of proteins A, G, and L, each of which bind to specific domains on an antibody/fragment, or the use of specific tags (e.g., histidine and biotin) attached to antibodies or antigens are also described.

摘要

本章介绍了肽抗体(如IgG、IgY、单链抗体和Fab片段)的纯化方法。亲和色谱纯化是一种非常方便且有效的抗体纯化策略,它基于特异性结合来分离肽抗体,即抗体与固定有其特异性配体的柱子结合,随后洗脱纯化的抗体。此外,还描述了基于使用蛋白A、G和L(它们分别与抗体/片段上的特定结构域结合)的纯化方法,或使用连接到抗体或抗原上的特异性标签(如组氨酸和生物素)的应用。

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