Antibody purification: ion-exchange chromatography.
Antibody purification: ion-exchange chromatography.
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DOI：
10.1007/978-1-59745-324-0_4
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发表时间：
2010
影响因子：
--
通讯作者：
A. Grodzki;E. Berenstein
中科院分区：
文献类型：
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作者：
A. Grodzki;E. Berenstein
离子交换层析技术是本章的重点，它们展示了这种方法在纯化蛋白质和单抗方面的力量。这项技术非常强大，可以分离净电荷有微小差异的生物分子，例如，两个蛋白质分子只有一个带电氨基酸的差异。鉴于蛋白质的两性特性，溶液的pH值在确定所用离子交换剂的类型时很重要。免疫球蛋白虽然可以通过阳离子交换层析或阴离子交换层析来纯化，但最常见的是用DEAE树脂进行阴离子交换来纯化。本文详细介绍了用DEAE柱从血清中纯化兔免疫球蛋白的方法，以及用FPLC从腹水中纯化免疫球蛋白的方法，从纯化的浓缩蛋白的负载到洗脱。
Ion exchange chromatography techniques are the focus of this chapter and they showcase the power of this method for the purification of proteins and monoclonal antibodies. The technique is powerful and can separate biomolecules that have minor differences in their net charge, e.g., two protein molecules differing by a single charged amino acid. Given the amphoteric character of proteins the pH of the solution is important in the determination of the type of ion exchanger used. Immunoglobulins, although they can be purified by either cation or anion exchange chromatography, are most frequently purified by anion exchange with DEAE resins. The purification of the rabbit IgG fraction from serum using a DEAE column is detailed as well as the purification of IgG from ascitic fluid using FPLC, from loading to elution of the purified and concentrated protein.