Validating microvesicle biomarkers for vascular disease (lacunar infarction) prognosis

• Main Author: Ng, Wei Ling
• Other Authors: Sze Siu Kwan
• Format: Final Year Project
• Language: English
• Published: 2014
• Subjects: DRNTU::Science
• Online Access: http://hdl.handle.net/10356/61712
• Institution: Nanyang Technological University

Despite of having a good rate of recovery, LACI patients show poor mid- to long-term prognosis, either recurrence of vascular events or decline in cognitive functions. Microvesicles, which are membrane bound organelles released from the plasma membrane by many types of cells (Dittmar and Zanker 2011), were used as the source of biomarker. Downstream proteomics profiling was performed to discover the biomarkers for LACI prognosis in these plasma microvesicles. This study is focused on validating the myelin basic protein (MBP) as a biomarker for LACI prognosis in different groups of LACI subjects by developing a new and simple protocol to isolate microvesicles in human plasma using smaller plasma volumes. Two protocols were performed. The ultracentrifugation protocol was performed and the results were analyzed by using western blot and LC-MS/MS. MBP signal was detected in individual subject. However, no MBP and very weak CD9 markers were detected when the experiment was repeated with less plasma sample. The new protocol, which was much simpler and time-saving, was performed with even lesser plasma samples, yet strong signals of MBP and CD9 were detected. Therefore, we can conclude that we establish a highly efficient and effective new protocol to isolate the microvesicle biomarkers.