Department of Pure and Applied Biochemistry, Centre for Chemistry and Chemical Engineering, Lund University, Lund, Sweden.
J Chromatogr A. 2010 Jun 4;1217(23):3635-41. doi: 10.1016/j.chroma.2010.03.050. Epub 2010 Apr 1.
In this paper, we present a new modular approach to immobilize boronic acid ligands that can offer effective separation of glycoproteins. A new "clickable" boronic acid ligand was synthesized by introducing a terminal acetylene group into commercially available 3-aminophenyl boronic acid. The clickable ligand, 3-(prop-2-ynyloxycarbonylamino)phenylboronic acid (2) could be easily coupled to azide-functionalized hydrophilic Sepharose using Cu(I)-catalyzed 1,3-dipolar cycloaddition reaction under mild condition. Compared to other boronic acid affinity gels, the new affinity gel displayed superior effectiveness in separating model glycoproteins (ovalbumin and RNase B) from closely related bovine serum albumin and RNase A in the presence of crude Escherichia coli proteins. Because of the simplicity of the immobilization through "click chemistry", the new ligand 2 is expected to not only offer improved glycoprotein separation in other formats, but also act as a useful building block to develop new chemical sensors for analysis of other glycan compounds.
在本文中，我们提出了一种新的固定硼酸配体的模块化方法，该方法可以有效分离糖蛋白。通过在市售的 3-氨基苯硼酸中引入末端乙炔基，合成了一种新的“点击化学”硼酸配体 3-(丙-2-炔氧基羰基氨基)苯硼酸（2）。在温和条件下，该点击化学配体 2 可以很容易地与叠氮基功能化的亲水 Sepharose 通过 Cu(I) 催化的 1,3-偶极环加成反应偶联。与其他硼酸亲和凝胶相比，新的亲和凝胶在存在粗大肠杆菌蛋白的情况下，从牛血清白蛋白和 RNase A 等密切相关的蛋白质中分离模型糖蛋白（卵清蛋白和 RNase B）的效果更好。由于通过“点击化学”进行固定的简单性，新配体 2 不仅有望在其他形式中提供改进的糖蛋白分离，而且还可以作为开发用于分析其他糖化合物的新型化学传感器的有用构建块。
