ABSTRACT
This chapter explores the chromatographic properties of a silica-based weak cation-exchange high-performance liquid chromatography (HPLC) column using acidic volatile buffer solutions. Cation-exchange high-performance liquid chromatography (CEC) was carried out on equipment from Millipore-Waters comprising a Model 680 automated gradient controller supplied with two 510 solvent pumps and a 481 variable wavelength detector linked to a BD41 chart recorder from Kipp. In ion-exchange chromatography, the choice of the type of matrix depends on the isoelectric point and the stability within a certain pH range of the proteins to be separated. The proteins were divided into three groups representing the highly basic proteins, the acidic proteins, and an intermediate class of proteins, called the neutral proteins. The pH plays a more important role than ionic strength in the separation of these acidic proteins. The conditions determined for optimal separation of proteins on an analytical/semipreparative column can probably also be applied to preparative CM-2-SW cation exchangers.
