Capillary-based separation microtechnologies for pattern profiling of phosphoproteins involved in T cell signaling

Current methods of determining pathogen exposure are bound by the proliferation of the pathogens within the bloodstream, by antibody production, or by the appearance of symptoms as the disease progresses. It is therefore advantageous in treatment, or containment, to have a technique that can detect exposure at an earlier stage. Because protein phosphorylation is an integral part of cell signaling, and one of the earliest indicators of immune system activation, phosphoproteins are an attractive target. We have used sensitive micro-analytical separation techniques, such as capillary gel electrophoresis (CGE) coupled to laser induced fluorescence detection, to probe differences in phosphoprotein populations extracted from immune cells. Using both general and specific modes of activation, we have consistently been able to differentiate between profiles or ´fingerprints´ from resting and activated T cells. Further experiments are underway to determine the robustness of the method. Development of this application may lead to a tool for rapid determination of immune system activation, which can be beneficial to the medical and biological research communities