The Use of 2D-LC-MS/MS in Disease Characterization and Global Proteomics

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2006-06

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The Ohio State University

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Abstract

Proteins occupy more than 50% of the dry weight of the average human cell. The proteins present in a given cell at a given time reflect the function and specialization of that cell. In addition to the specialization of cells within the body leading to various normal protein states, protein ‘sub-states’ can be induced by various conditions such as infections or cancer [9,21]. These disease-induced protein sub-states result in both qualitative and quantitative differences in both ribosomal production of proteins and posttranslational modification of proteins [2]. Each disease-induced sub-state therefore has its own protein profile or signature. For diseases such as cancer, which can easily progress undetected and whose manifestations have grave consequences, statistical comparisons of a patient’s protein profile with many cancer-state protein profiles can be made for early detection. Two-dimensional-liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) holds much promise in the analysis of such diseaseinduced protein profiles. 2D-LC-MS/MS uses two means of separation prior to online mass analysis, whereas one-dimensional (1D) methods use only one means of separation. The increased separation of a 2D system allows for much more complete mass analysis of proteins and results in the identification and characterization of many more proteins and modifications than with a 1D system [15]. By comparing patients’ protein profiles with a comprehensive proteomic database, 2D-LC-MS/MS could serve as the critical step in an efficient early-detection method for diseases such as cancer. Before protein profiles can be accurately compared and linked with diseases, a standardized technique must be used, and mechanisms that account for experimental variations must be implemented. The 2DLC- MS/MS technique proposed here is both very useful and can be standardized easily.

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Cancer, bioinformatics, proteomics, LC-MS

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