Hatch Duane M, Weiss Alison A, Kale Ramesh R, Iyer Suri S
Department of Chemistry, University of Cincinnati, Cincinnati, Ohio 45221-0172, USA.
Chembiochem. 2008 Oct 13;9(15):2433-42. doi: 10.1002/cbic.200800188.
Glycans cover the surface of all mammalian cells. Several toxins and pathogens use these glycans to bind and infect the cell. Using a versatile modular synthetic strategy, we have developed biotinylated bi- and tetraantennary glycoconjugates to capture and detect E. coli and compared the capturing ability of these molecules to commercial polyclonal antibodies. Magnetic beads were coated with biotinylated glycoconjugate or antibody, and these beads were used to capture, isolate, and quantify bacterial recovery by using a luminescence assay. The glycoconjugate-coated magnetic beads outperformed antibody-coated magnetic beads in sensitivity and selectivity when compared under identical experimental conditions. Glycoconjugates could capture Escherichia coli from stagnant water, and the ability of a panel of glycoconjugates to capture a selection of pathogenic bacteria was also evaluated. To the best of our knowledge, this study represents the first comprehensive study that compares synthetic glycoconjugates and antibodies for E. coli detection. The glycoconjugates are also very stable and inexpensive. The results presented here are expected to lead to an increased interest in developing glycoconjugate-based high affinity reagents for diagnostics.
聚糖覆盖所有哺乳动物细胞的表面。几种毒素和病原体利用这些聚糖来结合并感染细胞。我们采用通用的模块化合成策略，开发了生物素化的二天线和四天线糖缀合物，用于捕获和检测大肠杆菌，并将这些分子的捕获能力与市售多克隆抗体进行比较。将磁珠用生物素化糖缀合物或抗体包被，然后利用发光测定法，用这些磁珠捕获、分离并定量细菌回收率。在相同实验条件下比较时，糖缀合物包被的磁珠在灵敏度和选择性方面优于抗体包被的磁珠。糖缀合物能够从死水中捕获大肠杆菌，还评估了一组糖缀合物捕获多种病原菌的能力。据我们所知，本研究是第一项比较合成糖缀合物和抗体用于检测大肠杆菌的综合性研究。这些糖缀合物也非常稳定且价格低廉。此处呈现的结果有望引发人们对开发基于糖缀合物的高亲和力诊断试剂的更大兴趣。
