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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。
In order to construct a simple and efficient method for the isolation and purification of pathogenic microorganisms, a bimetallic magnetic nanoparticle Au-Fe3O4 was prepared. Then, the surface of the Au component was modified with lactose (Lac) which could specifically bind lectins and pathogenic microorganisms. The effect of Lac-Au-Fe3O4 and RCA120 on the separation efficiency of RCA120 was also studied. The results show that: 1) Au- Fe3O4 nanoparticles were uniform in size (8 +14 nm), dumbbell-like, and Au and Fe3O4 were located at both ends of the dumbbell respectively. 2) Lac-Au-Fe3O4 had good water solubility and magnetic properties, And fully combined with RCA120; 3) Under the applied magnetic field, the separation efficiency of RCA120 in aqueous solution by Lac-Au-Fe3O4 is about 70%, which can effectively separate the trace sample (1nmol / L). Through this experiment, the author has a more in-depth understanding of the preparation of glycosylated magnetic nanoparticles and their reaction with lectin, which laid the foundation for the further development of a simple and rapid method for the separation of pathogenic microorganisms.