糖基化磁性纳米粒子的制备及其对凝集素的富集分离

Purification of lectin using magnetic glyco-nanoparticles

为了构建简单高效的病原微生物分离纯化方法,首先制备了双金属磁性纳米粒子Au-Fe3O4,然后在Au组份的表面修饰了可特异性结合凝集素及病原微生物的的乳糖基(Lac),获得了糖基化磁性纳米粒子Lac-Au-Fe3O4,并进一步研究了Lac-Au-Fe3O4与蓖麻凝集素(RCA120)之间的反应及其对RCA120分离效率的影响,结果表明:1)Au-Fe3O4纳米粒子尺寸均一(8+14nm),呈哑铃状,Au和Fe3O4组份分别位于哑铃的两端;2)Lac-Au-Fe3O4具有良好的水溶性和磁学性质,在30min之内可特异性地与RCA120充分结合;3)在外加磁场作用下,Lac-Au-Fe3O4对水溶液中RCA120的分离效率约为70%,可有效分离微量样品(1nmol/L)。通过本实验,笔者对糖基化磁性纳米粒子的制备及其与凝集素的反应有了较为深入的了解,为进一步发展简单、快速的病原微生物分离方法奠定了基础。

Magnetic Au-Fe3O4 nanoparticles （NPs） were prepared and their surfaces were modified with a carbohydrate,lactose（Lac） ,which have binding capability to a lectin RCA120. Characterizations of the resulting glyco- NPs（Lac-Au-Fe3O4） reveal that they have uniform dumbbell-like shapes（8 ＋ 14 nm） with Lac residues coupled to the Au components. The Lac-Au-Fe3O4 NPs showed good solubility allowing a fast and full interaction with RCA120 in an aqueous solution within 30 minutes. By applying a magnetic field,RCA120 was further isolated from the solution with the highest rate of separation at approximately 70%. Such purification could be achieved even when the concentration of RCA120 was as low as 1 nmol/L. These results demonstrated that the potentials of carbohydrate-functionalized Au- Fe3O4 NPs in rapid and fast purification of lectins,which are commonly found on the surface of many pathogens, thereby providing a baseline for the further studies on microbial purifications by this new promising material.