The explosive growth in biological, pharmaceutical, and human health screening technologies has challenged the analytical sciences to create methodologies for high-sample throughput. Of the many possible methodologies, capillary electrophoresis (CE) has an unrecognized potential to serve as a platform for a highly multiplexed analysis system that allows for extremely high sample throughput. However, CE traditionally suffers from unacceptably high sample-to-sample and capillary-to-capillary variations in elution profiles and injection volumes. This variability is one of the main impediments to the use of CE in routine analysis. Furthermore, today's multiplexed CE systems are based only on fluorescence read-out concepts, which are ineffective in detecting -90 percent of the likely target compounds. This project describes the design fabrication, and implementation of a 96-capillary array CE system that 1) employs an absorbance-based read-out methodology and 2) utilizes the near-continuous measurement, collection, and application of the current flow at each individual capillary. The former significantly expands the scope of the system. The latter uses the collected current for self-normalization of the electropherogram through application of the migration index, which can yield precisions that compare favorably with that of high performance liquid chromatography. Success in Phase I will lead to a Phase II program focused on optimizing the system and methods to test biological fluids to perform disease diagnosis and/or prognosis as well as combinatorial drug synthesis and activity screening. Phase II would accumulate in actual field tests being performed and compared to traditional, accepted methods.
The ultimate goal of the project for commercial applications is the creation of a packaged system that has proven capabilities for solving problems in the health-related applications (e.g., cancer gene detection, multation analysis, forensic genotyping and drug efficacy) and other areas demanding reliable, high throughput analysis.
| Grossmann, A H; Kolibaba, K S; Willis, S G et al. (2004) Catalytic domains of tyrosine kinases determine the phosphorylation sites within c-Cbl. FEBS Lett 577:555-62 |
