Biomedical applications of two-dimensional electrophoresis using immobilized pH gradients: Current status

There is currently much interest, as we enter the postgenome era, in studyi ng gene expression at the protein level. Two-dimensional electrophoresis (2 -DE) using immobilized pH gradients (IPG), coupled with mass spectrometry ( MS), is currently the most widely utilized approach for the analysis of who le tissue proteins. The methodology for IPG-based 2-DE, since the introduct ion of the technique in the 1980s, is reviewed. In its present form the IPG methodology is mostly useful as a research tool. In general, high reproduc ibility and high resolution have been achieved. However, the lack of substa ntial automation and the limited sensitivity of the current overall methodo logy continue to represent drawbacks for biomedical applications. Further d evelopments to increase throughput and to reduce sample requirement would s ubstantially benefit the application of IPG-based 2-DE to biomedicine and w ould enhance the prospects for introducing the methodology into the clinica l laboratory.