Moosmann Anna, Müller Egbert, Böttinger Heiner
Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, Stuttgart, 70569, Germany,
Methods Mol Biol. 2014;1129:527-38. doi: 10.1007/978-1-62703-977-2_37.
PEGylation is a common and highly accepted possibility for half-life prolongation of proteins by increasing the hydrodynamic size. The chromatographic purification of PEGylated protein, using PEG (poly-ethylene glycol) of different PEG chain lengths, with the example of lysozyme and a scFv, is described in detail here, and helpful suggestions for the purification of other PEGylated proteins are listed. The relevant characterization methods for PEGylated proteins, important for the successful purification, are also described. The purification starts with a CEX (cation exchange) chromatography leading to about 95 % purity for polishing HIC (hydrophobic interaction chromatography) is described.
聚乙二醇化是通过增加流体力学尺寸来延长蛋白质半衰期的一种常见且被广泛接受的方法。本文详细描述了使用不同聚乙二醇链长的聚乙二醇（PEG）对溶菌酶和单链抗体片段（scFv）进行聚乙二醇化蛋白质的色谱纯化，并列出了纯化其他聚乙二醇化蛋白质的有用建议。还描述了聚乙二醇化蛋白质的相关表征方法，这些方法对成功纯化很重要。纯化过程首先通过阳离子交换（CEX）色谱，然后进行疏水相互作用色谱（HIC）以达到约95%的纯度进行精制。
