Investigation on the Biomarkers of Autoimmune Disease by Mass Spectrometry and Surface Plasmon Resonance

• Main Authors: Yu-Ren Chen, 陳昱任
• Other Authors: Wen-Yih Chen
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/02415616880114391048

碩士 === 國立中央大學 === 化學工程與材料工程學系 === 101 === Biomarker is a biologic analytical method to measure the normal physiological reaction, to understand the pathology, and drug safety. Besides, biomarker can be a characteristic for decision making in clinical diagnosis. The applications of biomarker include etiopathology understanding and the effect of follow-up care and the drug discovery. In this study, we investigated the biomarkers to rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) detected by mass spectrometry (MS) and surface plasmon resonance (SPR), respectively. There are two topics involved in this research. First one is to obtain the biomarkers for anti-tumor necrosis factor (anti-TNF) and anti-CD20 which are the possible treatment to RA using mass spectrum. In this topic, we combined MS with Fe3O4/TiO2 nano-particle to search for the new biomarkers applied to different biologics from patient serum. The other topic is a cohort study on detecting anti-double strained deoxyribonucleic acid antibody (anti-ds DNA antibody) from the serum of SLE patient. We used gold chip immobilized with dsDNA to detect anti-dsDNA antibody by our lab-owned SPR, and compared it with enzyme-linked immunosorbent assay (ELISA) on the sensitivity and specificity. From the results of first topic, we successfully enhanced the detection signal performance of MS by Fe3O4/TiO2 nano-particle and found out two biomarkers with molecular weight of 3.89 kDa and 7.77 kDa. In specific, from the result treated with Enbrel, the specificity using 3.89 kDa or 7.77 kDa biomarker approximates to 75%. And the sensitivity using 3.89 kDa and 7.77 kDa biomarkers approximate to 83% and 79%, respectively. Besides, from the result treated with Humira, the sensitivity and specificity using 7.77 kDa are 70% and 75%, respectively. The second topic is a cohort study, that is, a detection platform establishment to detect anti-ds DNA antibody with the sensor chip which synthesized by mixed self-assembling membrane (SAM) technique and EDC/sulfo-NHS surface modification. We have collected the information from thirteen SLE patients, obtained the serum samples from 24 patients in different time and the controlling variables from 4 normal people. From the receiver operating characteristic (ROC) curve and the p value measurement, we found that the recognition efficiency of dsDNA to anti-dsDNA antibody from SPR detection is better than that of from ELISA, so does the diagnose value. In general, we can successfully establish a detection platform with high sensitivity and specificity to detect the anti-dsDNA from serum and provide the guidance for early diagnosis in the clinical medicine, which is a significant achievement in nowadays translational medicine.