Development of a Capillary Electrophoresis (CE)-based Integrated Analytical Platform for Low Abundance Proteins

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Proteomics, a large-scale study of proteins in biological systems, led to a paradigm shift in the understanding of human physiology and drug discovery advances. Mass spectrometry (MS) is an essential tool in proteomics. Advancements in MS over the past decade have resulted in dramatic improvements in speed, reliability and most importantly, sensitivity. Sensitivity is vital in the discovery of low abundance proteins that can be crucial drug targets. While today's MS technology is capable of detecting very small quantities of proteins, most of the currently existing sample preparation procedures require much larger amount of proteins. To fully utilize the power of advanced MS, handling of small quantities of proteins in very small volumes is essential. The research proposed herein focuses on the development of a small-volume, sub-microliter, sample-handling platform based on an advanced analytical technique called capillary electrophoresis (CE). Yeung has previously demonstrated the use of CE to perform a number of sample preparation reactions for minuscule quantities of proteins. In this application, he proposes the integration of these individual reactions using magnetic microbeads for analyte transport at minuscule quantities. Such an integrated analytical platform will allow the entire multiple-step sample preparation to be conducted in sequence at a miniaturized level. The outcome is expected to enable ultrasensitive protein analysis when sample quantity is severely limited. To illustrate its benefits to society, the developed methodology will be applied in two areas. The first one is analysis of the protein film formed on tooth surfaces. The composition of this film directly affects the oral microbial activities related to tooth decay and periodontal infection. The second application focuses on protein kinase CK2, which is commonly found at elevated levels in cancer cells. The proposed new methods will be applied to study the inhibiting effects of CK2 in programmed cell death; i.e., CK2's role in cancer cells. Both of these studies, enabled by the newly developed techniques, will lead to immediate advances in prevention and treatment of these diseases.