Lipidomic profiling of bacterial and cancer cells using MALDI-MSand Raman spectroscopy- a nanomaterial based analytical approach

• Main Authors: Pei-yang Hua, 華沛揚
• Other Authors: Hui-Fen Wu
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/8592er

碩士 === 國立中山大學 === 化學系研究所 === 101 === Lipidomics can be defined as the characterization of lipid molecules and their interactions in biological systems i.e. a system-level analysis for lipidome. Lipids are an integral part of biomolecules. In addition to this, they also are a part of cell membranes, store energy, participate in the expression of proteins and are involved in gene regulation for biochemical actions. The Nobel Prize in Chemistry in 2002, Koichi Tanaka and John Fenn was awarded to the development of MALDI and ESI techniques. Both these techniques have been applied to biomolecule analysis and paved a doorway for interdisciplinary research involving, genomics, proteomics, lipidomics. With the introduction of nanotechnology, the detection sensitivity and the ability to analyze real samples have been made possible through nanomaterial based mass spectrometry. In the first application, lipids from cancer and cancer stem cells where extracted in organic solvent systems and graphene has been used as matrix for the analysis of the cancer lipids using MALDI-MS analysis. For the first time the application of graphene-assisted laser desorption ionization mass spectrometry (GALDI-MS) for lipidomics profiling of cancer and cancer stem cells has been demonstrated. We have also used this matrix for the successful differentiation between normal breast cells and breast cancer cells and cancer stem cells. This approach can be used for the discovery of potential biomarkers in biomedicine based on lipidomic profiling. In the second application, we reported the successful inhouse synthesis of ZnO nanoparticle (NPs), quantum dots (QDs) and nanorods (NRs). All these three ZnO nanomaterials were tested for their toxicity/compatibilty towards Pseudomonas aeruginosa and Staphylococcus aureus. The growth pattern of both these bacterial pathogens in the presence of the ZnO nanomaterials and the subsequent lipidomic changes were assessed using MALDI-MS. In the third application, for the first time, the use of CuFeO2@chitosan and Fe3O4@chitosan nanomagnets for affinity based separation and enrichment of trace levels of endotoxin and direct detection using MALDI-MS has been reported. The results showed that the CuFeO2@chitosan nanomagnets appeared to be more effective than Fe3O4@chitosan nanomagnets. Consequently, this approach proposes a novel MALDI-MS platform that can be further applied for biomedicine/clinical applications for rapid, direct and effective detection of bacterial infections. In the fourth application, rapid differentiation of gram negative and gram positive bacteria based on nanoparticle enabled fatty acid profiling using nano-Raman and nano-MALDI-MS. The results showed that Au/Pt NPs in the ratio of 1:1 combined with 9-AA matrix could lead to distinct lipid profiling for differentiation between gram positive and gram negative bacteria using nano-MALDI-MS. Also, in case of nano-Raman using Au NPs/Pt NCs as solid Raman substrates, specific peaks unique to specific bacteria were identified.