Open Competition 3 - Biotechnology

Multiplexed Capillary Electrophoresis for Comprehensive Peptide Mapping

Sponsor: CombiSep, Inc.

• Project Performance Period: 2/1/2002 - 1/31/2005
• Total project (est.): $3,175,105.00
• Requested ATP funds: $1,992,558.00

The recent completion of the "rough draft" chemical sequence of the human genome is the first step in obtaining genetic information to help elucidate the origins of diseases and biological functions. The next step, which is more complex, is to characterize the structure and function of gene products -- proteins -- which direct biological activities. Recent studies suggest that each gene produces about 10 different proteins. CombiSep Inc. plans to develop and demonstrate technologies for a rapid, low-cost protein characterization scheme based on peptide mapping. Peptides are protein metabolites that reveal overall system function. Peptide mapping -- which involves cleaving a sample protein by enzymes or chemical digestion and identifying the resulting molecules -- allows rapid analysis with simple instrumentation and will definitively identify most proteins, even revealing mutations and other rare modifications. In the three-year project, the company will extend the capabilities of ultraviolet-absorbance-based capillary electrophoresis (CE) technology developed at Iowa State University (Ames, Iowa), which will be subcontracted to provide engineering services. CE is a method for separating molecules in a charged fluid or gel that, among other advantages, can be multiplexed and automated so that many experiments are performed in parallel. Researchers will develop a validated set of separation conditions so that peptides can be identified based on the time it takes to migrate through the CE system, combined with other information. The company will develop reagents, methods for normalizing migration times and sample injection volumes, and a miniaturized two-dimensional channel electrophoretic system expected to provide higher resolution and sensitivity than conventional methods. A standardized, computer-searchable database and search engines will be developed to match CE results with known peptides using pattern recognition. ATP support is needed because CombiSep is a startup company, and venture capital is scarce in Iowa. If successfully developed and commercialized, the new technology will enable screening of protein profiles for purposes of clinical diagnosis, biotechnology production, and studies in functional genomics. It will offer high accuracy and reliability, 100 to 400 times the speed of existing protein analysis instruments, and a 30-fold reduction in instrument cost. The peptide database will significantly enhance U.S. competitiveness in the pharmaceutical and medical fields and open up new markets for CE instrumentation.

For project information:

Shelley J. Coldiron, (515) 294-7135
shelley.coldiron@combisep.com

ATP Project Manager

Thomas Wiggins, (301) 975-5416
thomas.wiggins@nist.gov