Department of Chemical Engineering, McMaster University, Hamilton, Ontario, Canada.
J Pharm Sci. 2010 Aug;99(8):3326-33. doi: 10.1002/jps.22103.
N-terminus-specific PEGylation was used to produce mono-PEGylated lysozyme. However, some di- and tri-PEGylated proteins were also produced due to side chain reaction. The reaction products were characterized by chromatographic and electrophoretic methods. Commercial cation exchange membrane Sartobind S was used for chromatographic purification of PEGylated lysozyme, the basis of separation being the shielding of protein charge by PEG. Using the membrane chromatographic method, lysozyme and mono-, di-, and tri-PEGylated lysozyme could be resolved into separate peaks. Increasing the superficial velocity during chromatographic separation from 24 cm/h to 240 cm/h increased both protein binding capacity and resolution due to enhancement of protein mass transfer coefficient.
采用 N 端特异性聚乙二醇化方法制备单聚乙二醇化溶菌酶。然而，由于侧链反应，也产生了一些二聚体和三聚体的聚乙二醇化蛋白。通过色谱和电泳方法对反应产物进行了表征。商业阳离子交换膜 Sartobind S 用于聚乙二醇化溶菌酶的色谱纯化，分离的基础是聚乙二醇对蛋白质电荷的屏蔽。使用膜色谱法，可以将溶菌酶和单聚、二聚和三聚体的聚乙二醇化溶菌酶分别解析成单独的峰。在色谱分离过程中，将表面速度从 24cm/h 增加到 240cm/h，由于蛋白质传质系数的提高，同时增加了蛋白质的结合容量和分辨率。
