製備高磁性四氧化三鐵奈米材料及研究其在C反應蛋白分離上的應用

Preparation of High Magnetization Fe3O4 Nanomaterials and Investigation of Their Application in Separation of C-reactive Protein

賴韋翔(Wei-Hsiang Lai)

指導教授 ： 陳家俊

國立臺灣師範大學/理學院/化學系/碩士(2010年)

摘要

在本文中我們合成一致性都相當高的四氧化三鐵磁性奈米粒子，藉由反應條件不同來調控粒徑大小的差異，探討粒徑大小對材料磁性的強度的關係。利用配位體置換反應的表面修飾改良表面二氧化矽磁力遮蔽問題，合成出較高磁性的磁性奈米粒子，再由於我們在其表面修飾上胺基，利用自由胺基容易繼續修飾其他分子的特色，最終形成表面修飾上C反應蛋白的1級抗體的磁性奈米複合材料，並利用多胺化合物提高表面自由胺基的量，討論提升自由胺基的量對表面修飾1級抗體的影響，而在本文內我們藉由一系列的實驗方法佐證最終複合材料表面的1級抗體具活性，並利用磁性奈米材料的特性以及抗原抗體間的高專一性、高結合力，自動化且有效率的分離出血清中的C反應抗原蛋白。
　　另外我們利用磁性奈米材料的特性發展出一套自動化分離純化C反應蛋白的流程，我們將功能性磁性奈米複合粒子直接加到人類血清中去分離純化C反應抗原蛋白，在施以外加磁場的情況下，不需要離心的動作即可完成多組血清樣品分離純化C反應蛋白抗原，最後可以再配合MALDI-TOF Mass做質譜分析，並利用磁性奈米材料取代傳統三明治ELISA免疫分析方法中的底材，利用材性奈米材料的特性，分離出C反應蛋白，藉由標準曲線，推算出血清中C反應蛋白的濃度。

關鍵字

四氧化三鐵 ； 奈米材料 ； 磁性

並列摘要

In this article, we synthesized magnetic Fe3O4 nanoparticles(NPs) of high magnetic response. We adjusted the differences in particle size through different reaction conditions, and also explored the impact of particle size on the magnetic intensity. By modifying and improving silica magnetic shielding problem on the surface with ligand displacement reaction, it was definite to synthesize Fe3O4 NPs with higher magnetism. Because the surface was modified with amino ligands, the NPs were able to conjugated with C-reactive protein(CRP) antibody easily to form functional magnetic NPs(Fe3O4@ab-CRP NPs). Furthermore, the amount of primary amino ligands on the surface were increased with antibody surface conjugation. Also, we proved that the primary antibody on the surface of Fe3O4@ab-CRP NPs still exhibited bio-activity.

　　Besides, we utilized the features of the high specificity and high bonding capacity between antigen and antibody to separate CRP antigen protein. We have developed an automated process for separating purified CRP using the features and magnetism of Fe3O4@ab-CRP NPs. By adding Fe3O4@ab-CRP NPs directly to human serum to separate and purify CRP while applying external magnetic field, it was possible to complete multiple separation of purified CRP separation on serum sample without undergoing centrifugal steps. Then through mass spectrum analysis using MALDI-TOF Mass and sandwich ELISA with functional magnetic NPs, both methods can finally separate the CRP, and calculate concentration through standard curve.

並列關鍵字

Fe3O4 ； Nanomaterials ； Magnetization

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