硼酸分子可在微鹼性環境與醣類結構上之雙醇基團(cis-diol)產生親和性交互作用，本研究選用胺基苯硼酸(Aminophenylboronic acid)為親和配體接附於奈米級的磁性微粒上，利用硼酸之親和特性，吸附分離醣蛋白，達到簡易操作、快速省時之目的。
首先以共沉澱法製備粒徑為20-30 nm且具超順磁性的四氧化三鐵(Fe3O4)顆粒，分兩種方式進行兩階段修飾，第一種方式是先以多巴胺(Dopamine)，第二種方式則是先以胺基矽烷aminopropyltriethoxysilane(APTES)修飾磁粒表面，分別可測得表面胺基含量為0.986 mmol/g DW與1.63 mmol/g DW，第二階段再利用戊二醛固定胺基苯硼酸於磁粒表面，分別可測得表面硼酸約有0.334 mmol/ g DW與0.516 mmol/g DW，所製備硼酸修飾後之磁粒仍具有磁性強度約68 emu/g。
此硼酸配體磁性微粒，在pH 9、50 mM HEPES buffer (20 mM Mg2+，10 mM NaCl)中有最佳吸附醣蛋白HRP的效果。多巴胺與矽化修飾硼酸配體磁粒對HRP之最大吸附量，由Langmuir恆溫吸附模式可分別算出約60.24與135.69 mg/g DW，但在使用 1 mg/ml HRP濃度下，實際吸附約為26與43 mg/g DW。再以200 mM山梨糖醇溶液脫附，可分別得到約2,000與10,000 U/g DW HRP活性。
硼酸配體磁性微粒可應用於多數的醣化蛋白純化，從血清中純化抗體(IgG)，以及雞蛋蛋清中Ovotransferrin、Ovalbumin、Ovoflavoprotien三種醣蛋白，吸附結果具有專一性，可有效純化濃縮醣蛋白；由SDS-PAGE電泳實驗分析，對醣蛋白的吸附量為矽化修飾硼酸配體磁粒＞商業Clontech硼酸膠體＞多巴胺修飾硼酸配體磁粒。

Taking advantages of the specific affinity interaction between 1,2-cis diol of glycoproteins and the boronic acid at mild alkaline conditon, we immobilized 3-aminophenylboronic acid as an affinity ligand on the surface modified magnetic nanoparticles for rapid and efficient enrichment of glycoproteins.
The magnetic nanoparticles (magnetite; Fe3O4) (MNP) were prepared by coprecipitation of Fe2+ and Fe3+ ions in the presence of base. The nanoparticles were about 20 - 30 nm in diameter and superparamagnetic as determined by TEM and SQUID, respectively. To obtain amino-functional surface on magnetic nanoparticles, we treated the the MNP with dopamine and 3-aminopropyltriethoxysilane (APTES) and obtained 0.986 and 1.63 mmol/g DW (dry weight) primary amino-group on the surface, respectively. Then ligand, aminophenylboronic acid was immobilized on magnetic nanoparticle via glutaraldehyde coupling. Approximately, 0.334 and 0.516 mmol/g DW of ligand was immobilized on the two different modified magentic nanoparticles, respectively. High saturated magnetism (68 emu/g) of the magnetic nanoparticles with boric acid functionalized on surface still can be obtained.
Horseradish peroxidase (HRP) was used as a model glycoprotein to study the affinity adsorption capacity of the as-prepared boronic acid-MNP. The adsorption was done in 50 mM HEPES buffer (20 mM Mg2+ and 10 mM NaCl) at pH 9 and room temperature. Langmuir adsorption model can be used to describe the adsorption isotherm and with maximum adsorption capacity Qmax of 60.34 and 135.60 mg/g DW for the dopamine and silane modified MNP, respectively. Based on HRP activity desorbed from the magnetic affinity absorbents by 200 mM sorbitol, the silane based MNP adsorbent shows 10,000 U/g DW which is much higher than that obtained from dopamine-based one (2,000 U/g DW).
The boronic acid surface functionalized magnetic nanoparticles also demonstrated selective capture capacity toward glycoproteins in serum and egg white as observed from SDS-PAGE analysis. As compared with Clontech’s commercial boronic acid resin, the boronic acid modified MNP prepared in this study shows the comparable affinity adsorption capacity as shown in the isolation of immunoglobulin from human serum and Ovotransferrin as well as ovalbumin from egg white.

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