College of Life Sciences, Northwest University, Xi'an, Shaanxi 710069, China.
Anal Bioanal Chem. 2010 Apr;396(8):3071-8. doi: 10.1007/s00216-010-3513-2. Epub 2010 Feb 19.
We introduce a novel combination of magnetic particles with hydrazine chemistry, dubbed as hydrazine-functionalized magnetic particles (HFMP) for isolation of glycopeptides. Four methods have been developed and compared for the production of HFMP by hydrazine modification of the surface of the carboxyl and epoxy-silanized magnetic particles, respectively. The evaluation of the capability and specificity of HFMP as well as the optimization of the coupling condition for capturing of glycoproteins were systematically investigated. The results showed that HFMP prepared by adipic dihydrazide functionalization from carboxyl-silanized magnetic particles (HFCA) displayed the maximum capture capacity and isolated efficiency for glycoprotein. When measured with glycoproteins, the capacity of the HFCA (1 g) for coupling bovine fetuin was 130 +/- 5.3 mg. The capability of this method was also confirmed by successful isolation of all formerly glycosylated peptides from standard glycoproteins and identification of their glycosylation sites, which demonstrated the feasibility of the HFCA as an alternative solid support for isolation of glycoproteins/glycopeptides.
我们介绍了一种新型的磁性粒子与联氨化学的组合，称为联氨功能化磁性粒子（HFMP），用于糖肽的分离。分别开发并比较了四种通过联氨修饰羧基和环氧硅烷化磁性粒子表面来生产 HFMP 的方法。系统研究了 HFMP 的性能和特异性以及用于捕获糖蛋白的偶联条件的优化。结果表明，由羧基硅烷化磁性粒子（HFCA）上的己二酰二肼功能化制备的 HFMP 显示出最大的糖蛋白捕获容量和分离效率。当用糖蛋白进行测量时，HFCA（1 g）用于偶联牛胎球蛋白的容量为 130 ± 5.3 mg。该方法的可行性还通过成功分离标准糖蛋白中的所有先前糖基化肽并鉴定其糖基化位点得到证实，这证明了 HFCA 作为糖蛋白/糖肽分离的替代固相载体的可行性。
