Kim Hyoung Sub, Hyun Ji Young, Park Seong-Hyun, Shin Injae
Center for Biofunctional Molecules, Department of Chemistry, Yonsei University Seoul 03722 Republic of Korea
RSC Adv. 2018 Apr 19;8(27):14898-14905. doi: 10.1039/c8ra01285g. eCollection 2018 Apr 18.
Pathogens infect hosts often through initial binding of their cell surface lectins to glycans expressed on the exterior of host cells. Thus, methods to evaluate the glycan-binding properties of pathogens are of great importance. Because of the multivalent nature of interactions of pathogens with glycans, the ability to assess the glycan density-dependent binding of pathogens is particularly important. In this study, we developed a facile technique to construct multivalent carbohydrate microarrays through immobilization of unmodified glycans on multivalent hydrazide-derivatized glass surfaces. This immobilization strategy does not require the use of multivalent glycoconjugates, which are typically prepared by using multistep sequences. The results of analysis of microarray images, obtained after incubation of multivalent glycan microarrays with cholera toxin B and pathogens such as uropathogenic and , show that the binding affinities of toxins and pathogens for glycans are highly glycan density-dependent. Specifically, toxins and pathogens bind to glycans more strongly as the valency of the glycans on the microarrays is increased from 1 to 4. It is anticipated that the newly developed immobilization method will be applicable to the preparation of multivalent carbohydrate microarrays that are employed to evaluate multivalent glycan binding properties of a variety of pathogens and toxins.
病原体通常通过其细胞表面凝集素与宿主细胞外表面表达的聚糖的初始结合来感染宿主。因此，评估病原体聚糖结合特性的方法非常重要。由于病原体与聚糖相互作用的多价性质，评估病原体聚糖密度依赖性结合的能力尤为重要。在本研究中，我们开发了一种简便的技术，通过将未修饰的聚糖固定在多价酰肼衍生化的玻璃表面上来构建多价碳水化合物微阵列。这种固定策略不需要使用通常通过多步序列制备的多价糖缀合物。在用霍乱毒素B和诸如尿路致病性大肠杆菌等病原体孵育多价聚糖微阵列后获得的微阵列图像分析结果表明，毒素和病原体对聚糖的结合亲和力高度依赖于聚糖密度。具体而言，随着微阵列上聚糖的价态从1增加到4，毒素和病原体与聚糖的结合更强。预计新开发的固定方法将适用于制备用于评估多种病原体和毒素的多价聚糖结合特性的多价碳水化合物微阵列。
