骨質疏鬆症是一種骨骼強?減弱，導致個人骨折危險性的疾病。檢測骨骼代謝的特定生化指標物質，為判斷骨質疏鬆症的重要依據。在尿液中骨質代謝指標之一的氮端胜?片段(NTx)抗原決定基為P1、P2、P3胜?，將雞隻多株抗體鍵結在經過表面官能基修飾的磁性?米?子上，再?用免疫性萃取反應搭配質譜分析技術與酵素連結免疫吸附法，檢測反應前、後待測溶液中殘餘胜?的濃度，以探討抗體與抗原結合的專一性。由基質輔助雷射脫附飛行時間質譜儀的分析檢測結果顯示，雞隻多株抗體對於P2胜?具有良好的結合專一性，對於P1、P3胜?則不具有顯著的結合性質。另外，由ELISA實驗結果發現，將P2抗體鍵結在孔盤上，能抓取P2-Ahx-FITC螢光胜?使得吸光值降低。最後，比較抗體鍵結在孔盤或磁性奈米粒子上的吸光值，以後者所測得的吸光值變化較大，有助於定量偵測。 Osteoporosis is a disease that leads to reduce bone strength and increase the risk of bone fracture. An important assessment for diagnosis of osteoporosis is the analysis of specific biomarkers of bone metabolism. In urine, one such biomarker found is the cross-linked N-telopeptide of type I collagen (NTx) epitope such as P1, P2, and P3 peptides. In this study, chicken polyclonal antibodies were bound on the modified surface of magnetic nanoparticles and then applied immunological reaction combining MALDI-TOF MS and ELISA techniques to detect the immune responses before and after the reaction. Peptide concentrations in the residual solutions were measured for the investigation of binding specificity between the antibodies and the antigens. The results of MALDI-TOF MS analysis showed chicken polyclonal antibodies had significantly binding specificity for P2 peptide. On the other hand, it did not have any binding property for P1 and P3 peptides. In addition, the ELISA results confirmed that the antibodies on the 96-wells plate could grab P2-Ahx-FITC fluorescence peptide and make the absorbance values decreased. Finally, comparing the absorbance values of P2 antibodies binding on the 96-wells plate or on the surface of magnetic nanoparticles, we found the later method showed larger absorbance change which could be used to improve quantitative analysis.
