Development of an LC-MS/MS-based system for the proteomic analysis of biological fluids and its application in assessing the effect of fuming oil exposure on the BALF proteome of rat

• Main Authors: Pang-Wei Chen, 陳邦維
• Other Authors: Pao-Chi Liao
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/94376694224764220381

碩士 === 國立成功大學 === 環境醫學研究所 === 93 === 　Biological fluids, such as serum, urine, cerebrospinal fluid (CSF), and lavage, are characteristics of different organ systems and body cavities and can be used for pathology monitoring. Especially, the proteins in body fluids are not only used for diagnosis but also contain important information related to pathology pathways. Proteomics refers to the systematic investigation of the proteins in a cell culture or a tissue, that is, the proteome, by separation, quantification, and identification of the complicated protein mixture. Today, the most popular analytical method for proteomic study is the combination of two-dimensional gel electrophoresis (2D-GE) and mass spectrometry (MS) protein identification (ID). However, 2D-GE process is time-consuming and highly labor-intensive. Alternative approaches that eliminating 2D-GE steps in proteomic study are desired and under intensive investigation. Recently, liquid chromatography tandem mass spectrometry (LC-MS/MS) has been used for proteome identification without tedious 2D-GE. We foresee that LC-MS/MS will be very useful to analyze biological fluids for the search of biomarkers related to various diseases. This study design and setup an LC-MS/MS-based system for proteomic analysis of biological fluids. This system can be used for the identification and quantification of proteomes in various biological fluids for biomarker discovery. The system should shorten the analysis time and reduce experimentation variations by automation. This analytical system has been used to study the changes in the proteome of the bronchoalveolar lavage fluid (BALF) of rats exposed directly to a fuming oil-releasing environment in a metal processing factory. The results revealed that 29 proteins exhibited significant changes after exposure. These proteins included surfactant-associated proteins (SP-A and SP-D), inflammatory proteins (complement component 3, immunoglobulins, lysozyme, etc.), growth factors (e.g. transforming growth factor alpha), calcium-binding proteins (calcyclin, calgranulin A, calreticulin, and calvasculin), and other proteins (e.g., cathepsin D, saposin, and intestinal trefoil factor). A large decrease in protein levels of SP-A and SP-D (0.24- and 0.38-fold, respectively) following exposure was observed. In contrast, protein levels of transforming growth factor alpha and calcium-binding proteins were significantly increased (4.46- and 1.4-1.8-fold, respectively). Due to the diverse functions of these proteins, the results might contribute to understand the mechanisms involved in lung disorders induced by oil mist exposure.