RPC GF (polishing steps)

Figure 2 Examples of purification strategies. * Major concentration steps; GF, gel filtration; IEX, ion exchange; HIC, hydrophobic interaction; RPC, reverse phase chromatography.

developed. It must be stressed that both media and instrumentation are continually being improved. Consequently, considerable time and effort can be saved by an understanding of the basic principles involved, and by analyzing any established protocol and looking for possible improvements. For example, a lengthy dialysis may be avoided by using the faster desalting method of gel filtration, and using an improved matrix may reduce a separation fivefold or more.

When problems do arise, it should always be remembered that no matter what the media or the protein extract, the basic principles governing the behavior and interaction of proteins between themselves and with other media remain the same.

See also: Affinity chromatography; Isoelectric focusing; SDS-polyacrylamide gel electrophoresis (SDS-PAGE).

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