Complex proteome prefractionation using microscale solution isoelectrofocusing.
The proteomes of mammalian cells, tissues and biologic fluids are complex and consist of proteins present over a wide dynamic range. Current protein profiling technologies do not have the capacity to overcome the sample complexity for comprehensive analysis of complex proteomes. A common strategy to substantially expand protein profiling capacities is sample prefractionation. A prefractionation method developed in the authors' laboratory, microscale solution isoelectrofocusing, has resulted in a commercial product, the ZOOM IEF Fractionator, which provides a simple and convenient method for high-resolution separation of complex proteomes based upon their isoelectric points. Complex human samples such as cancer cells and biologic fluids can be fractionated into well-resolved fractions with minimal cross-contamination of proteins between adjacent fractions. This review focuses on the application of microscale solution isoelectrofocusing prefractionation and subsequent downstream strategies in expanding protein profiling capacities and mining low-abundance proteins of complex proteomes.
The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA. tangh@wistar.org
Hsin-Yao
HY
Tang
David W
DW
Speicher
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods*; D008832:Microchemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012680:Sensitivity and Specificity
D020543:Proteome