?In this proposal we seek to demonstrate a new experimental approach to develop a new subtractive method to remove abundant proteins from complex human fluid samples. The ultimate goal is to develop a suite of methods or tools to facilitate the de novo discovery of a complete set of low-abundance potential protein biomarkers from which more targeted studies can be performed. A particular interest is in the discovery of potential biomarkers for cancers, and in the past we have studied lung cancer samples and references drawn from the same cohort. In this Phase I proposal, we seek to demonstrate the selective and specific removal of human serum albumin isoforms and dimers from human plasma. These serum albumins comprise half of the protein in human serum or plasma, and cause very real issues with both tandem mass spectrometry and difference gel electrophoresis approaches. The Phase II would extend to work to remove three orders of magnitude of abundant proteins, a 50 times increase from existing state-of-the-art methods. One key feature is that the conditions are chosen to minimize protein-protein interactions, which compromise the performance of existing methods. The simplified human fluid samples produced will be compatible with both 2D gel electrophoresis and liquid chromatography- tandem mass spectrometry approaches.
Public Health Relevance Statement: Public Health Relevance: Proteomics is a powerful approach that can be used to assess the state of a cell, tissue or organism. This proposal seeks to develop a new method to simplify complex protein mixtures derived from human fluids. It would allow a deeper mining of the proteome than existing state-of-the-art methods. The goal is to uncover very dilute proteins that indicate disease. It is intended as a research tool, and not one for the clinic, and the resuls would be used to create diagnostic tests using traditional methods. It would affect the fields of discovery of diagnostic markers to identify and development of pharmaceuticals that are used to treat disease.
NIH Spending Category: Biotechnology; Cancer
Project Terms: Adsorption; Affect; Affinity; Albumins; Antibodies; aptamer; Attention; Beds; Binding (Molecular Function); Biological Markers; Buffers; Cells; Chromatography; Clinic; cohort; combinatorial; Complement; Complex; cost effective; Data; Development; Diagnostic; Diagnostic tests; dimer; Disease; disulfide bond; DNA; Elements; Excision; expectation; gel electrophoresis; Goals; Human; interest; Libraries; Liquid Chromatography; Liquid substance; Malignant neoplasm of lung; Malignant Neoplasms; Marketing; Methods; Mining; MYO5A gene; Nucleic Acids; Organism; Performance; Pharmacologic Substance; Phase; Plasma; Polymethyl Methacrylate; Process; product development; protein complex; Protein Dynamics; Protein Isoforms; protein protein interaction; Proteins; Proteome; Proteomics; public health relevance; Research; Sampling; Serum; Serum Albumin; Solid; Solutions; Source; Specificity; surfactant; tandem mass spectrometry; Testing; Time; Tissues; tool; Two-Dimensional Gel Electrophoresis; Work
