The research presented here highlights projects focused on innovative means of analyzing complex biological samples. A common thread amongst the projects is to improve means for effectively and efficiently coupling of sample preparations and separations with mass spectrometry. The ability to accurately inventory and quantify the potentially 100s to 1000s of proteins that compose a given biological system often remains a challenge to bio-analytical science. Generally, the biggest obstacles to success are sample complexity coupled with a wide dynamic range of component concentration. Often, large-scale proteomic experiments must rely on separation science to distribute and concentrate the components prior to their sequentially detection. Without separation, the capability of mass spectrometers to distinguish the various components would be overwhelmed and rendered incomplete. Regardless, in many proteomic workflows, there remains a disconnection between optimization in sample preparations and separations and sample detection by mass spectrometry. The research outline here addresses this issue directly by targeting the construction of novel substrates (sample plates and membranes) designed to capture and concentrate biological analytes after separation, preserving the resolution gained, while acting as substrates for further sample processing (e.g., protein digestion, chemical modification). Additionally, many of sample substrates are uniquely designed to interface directly with the mass spectrometer.The applications for these novel sample plates and membranes have focused on linking protein separations or �top-down' proteomic schemes with mass spectrometry. Examples of these separations are gel-electrophoresis, isoelectric focusing, and protein chromatography. The �top-down� approach was targeted because it offers the best means for dealing with the wide dynamic range in concentration characteristic of biological samples (blood serum ~ 1012). Additionally, maintaining the protein structure helps preserve protein iso-form and glyco-form information; characteristics that may imply disease. In addition, innovative substrates designed to improve tissue-imaging analyses by MALDI mass spectrometry will also be presented.

Innovative strategies for coupling separations with MALDI mass spectrometry

246th National Meeting (2013)

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