Platform establishment to decrease non-specific plasma proteins/peptides applied with proteomic technology in tumor marker discovery.

• Main Authors: Sung-Yi Wei, 魏上宜
• Other Authors: Ann-Joy Cheng
• Format: Others
• Language: en_US
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/72966271185197305546

碩士 === 長庚大學 === 醫學生物技術研究所 === 94 === In current biomarker study, the MALDI-TOF mass spectrometry is a powerful proteomic tool to search and identify differential expressed plasma proteins between normal and cancer patients. Especially, the low molecular weight plasma proteome/peptideome has been focus of attempts to find novel markers. However, many limitations still exist. Recently study show that the differential peaks may be derived from endogenous proteolytic cleavage from abundant plasma proteins in response to general inflammation reactions. To overcome this problem, we establish a platform to decrease non-specific plasma proteins/peptides applied with proteomic technology aiming to facilitate tumor maker discovery. First, multiple antibodies based- affinity columns were used and shown successful depletion of six (Agilent) or twelve (GenWay) highest abundant proteins in plasma using PAGE analysis. Samples were then desalted and concentrated by micro-dialysis. Low molecular weight peptides were removal by C18-affinity magnetic beads as demonstrated by MALDI-TOF analysis. Finally, the optimal conditions for mass spectrometric analysis were examined. Proper dilution of samples was shown important step to obtain high resolution in mass analysis. Additionally, Samples were fractionated with various affinity magnetic beads (C8, C18, Cu, cation, anion) to reduce the complexity of plasma protein. In which, C18-bead was able to show a more complete plasma proteome/ peptideome profile, whereas Cu-bead was able to reflect more specific peaks. In the future, clinical sample study will be addressed with these considerations.