This report explores the issues faced by life scientists performing protein separation. To gain a comprehensive understanding of protein function and regulation, researchers must separate proteins, then identify and characterize the expressed proteins. 1-D or 2-D gel electrophoresis (2DGE) and occasionally single- or multi-dimensional liquid chromatography are the predominant techniques for separating proteins. The objective of this report is to explore the techniques and products used for protein separation. This vital information will help marketing professionals and product developers to better understand the users' level of satisfaction with the technologies available, the definition of quality from the users' perspective, and unmet needs of their customers and clients. 2-D gel electrophoresis allows protein scientists to separate either a whole cell or tissue protein extracts and then identify these proteins through peptide mass fingerprinting or via direct sequencing utilizing mass spectrometry (MS). Although 2DGE is the most widely used tool for separating proteins in expression proteomics, it is not without its limitations. Challenges faced when utilizing this technology are comigration of proteins, systematic exclusion of highly hydrophobic molecules, and problems with detecting very acidic, very basic, very small, very large, or low abundance proteins. To meet the demands of technology, companies are developing new innovations that appear to be inexpensive and reliable, generate high-resolution protein separation and yield good visual detection of subtle differences. Today gels are available as homogeneous, pre-cast products, which are combined with automated equipment and imaging analysis software for improved quantitative results and reduced workload. One example of a more advanced proteomic system is a 2-D gel electrophoresis system with common features such as an automated stainer, a 2-D analyzer, an automated gel exciser, an automated digester, and a robot for preMS sample preparation. Finally, as emerging technologies such as microcapillary electrophoresis, capillary electrochromatography and ultra-HPLC begin to replace 2-D gel electrophoresis, companies will need customer feedback in order to develop and maintain a competitive advantage in their product design and production. The final report is based on the opinions of a worldwide panel of research scientists. It provides you with the answers needed to better serve the protein science market and is available at a fraction of what it would cost to conduct a custom study similar in size and scope. Protein Separation is the third installment in our series - which details all aspects of proteomics - and specifically focuses on the well-established method of 2-D gel electrophoresis. Protein chemists widely use first-dimensional separation by isoelectric focusing and second-dimensional separation by molecular weight despite existing disadvantages or limitations. As interest in alternatives grow, companies involved in supporting protein research must understand life scientists' successes and frustrations with "state-of-the-art" technologies and how products must evolve to better meet the needs of this dynamic market. Report Highlights More than 500 researchers engaged in protein separation participated in this survey between September 4 and November 23, 2001. The report details findings for each and every question in the survey. Below is a glimpse of some of the findings derived from different questions: • The most preferred separation technique is one-dimensional SDS-polyacrylamide gel electrophoresis (PAGE) under reducing conditions. • Most respondents who use multiple separation techniques employ either single or multiple, tandem liquid chromatography approaches with or without subsequent mass spectrometry. • Over the next twelve months, usage of two cutting-edge approaches to separation technology - capillary gel electrophoresis and microfluidics "lab-on-a-chip" - in conjunction with mass spectrometry will significantly increase. • Respondents primarily use one-dimensional isoelectric focusing or one-dimensional SDS-PAGE for protein analysis/characterization experiments. • The protein scientists surveyed prefer to use two-dimensional gel electrophoresis for protein characterization and identification experiments but not for purifying their protein(s). • Over three-quarters of the scientists do not plan to use an integrated proteomics system that includes instrumentation specifically for protein separation. (Electronic copy also includes 1 print copy) | 
