Shang Jing, Piskarev Vladimir E, Xia Ming, Huang Pengwei, Jiang Xi, Likhosherstov Leonid M, Novikova Olga S, Newburg David S, Ratner Daniel M
Department of Bioengineering, University of Washington, 3720 15th Avenue NE, Seattle, WA 98195, USA.
Glycobiology. 2013 Dec;23(12):1491-8. doi: 10.1093/glycob/cwt077. Epub 2013 Sep 10.
Human milk glycans inhibit binding between norovirus and its host glycan receptor; such competitive inhibition by human milk glycans is associated with a reduced risk of infection. The relationship between the presence of specific structural motifs in the human milk glycan and its ability to inhibit binding by specific norovirus strains requires facile, accurate and miniaturized-binding assays. Toward this end, a high-throughput biosensor platform was developed based on surface plasmon resonance imaging (SPRi) of glycan microarrays. The SPRi was validated, and its utility was tested, by measuring binding specificities between defined human milk glycan epitopes and the capsids of two common norovirus strains, VA387 and Norwalk. Human milk oligosaccharide (HMOS)-based neoglycoconjugates, including chemically derived neoglycoproteins and oligosaccharide-glycine derivatives, were used to represent polyvalent glycoconjugates and monovalent oligosaccharides, respectively, in human milk. SPRi binding results established that the glycan motifs that bind norovirus capsids depend upon strain; VA387 capsid interacts with two neoglycoproteins, whereas Norwalk capsid binds to a different set of HMOS motifs in the form of both polyvalent neoglycoproteins and monovalent oligosaccharides. SPRi competitive binding assays further demonstrated that specific norovirus-binding glycans are able to inhibit norovirus capsid binding to their host receptors. A polyvalent neoglycoconjugate with clustered carbohydrate moieties is required for the inhibition of VA387 capsid binding to host receptor glycans, whereas both monovalent oligosaccharides and polyvalent neoglycoconjugates are able to inhibit Norwalk capsid binding to its host receptor. Binding of HMOS and HMOS-based neoglycoconjugates to norovirus capsids depends upon the specific strain characteristics, implying that HMOS and their polyvalent derivatives are potential anti-adhesive agents for norovirus prophylaxis.
人乳聚糖可抑制诺如病毒与其宿主聚糖受体之间的结合；人乳聚糖的这种竞争性抑制作用与感染风险降低有关。人乳聚糖中特定结构基序的存在与其抑制特定诺如病毒株结合的能力之间的关系需要简便、准确且小型化的结合测定方法。为此，基于聚糖微阵列的表面等离子体共振成像（SPRi）开发了一种高通量生物传感器平台。通过测量特定人乳聚糖表位与两种常见诺如病毒株VA387和诺沃克病毒衣壳之间的结合特异性，对SPRi进行了验证并测试了其效用。基于人乳寡糖（HMOS）的新糖缀合物，包括化学衍生的新糖蛋白和寡糖 - 甘氨酸衍生物，分别用于代表母乳中的多价糖缀合物和单价寡糖。SPRi结合结果表明，与诺如病毒衣壳结合的聚糖基序取决于病毒株；VA387衣壳与两种新糖蛋白相互作用，而诺沃克病毒衣壳则以多价新糖蛋白和单价寡糖的形式与另一组HMOS基序结合。SPRi竞争性结合试验进一步证明，特定的诺如病毒结合聚糖能够抑制诺如病毒衣壳与其宿主受体的结合。抑制VA387衣壳与宿主受体聚糖的结合需要具有聚集碳水化合物部分的多价新糖缀合物，而单价寡糖和多价新糖缀合物都能够抑制诺沃克病毒衣壳与其宿主受体的结合。HMOS和基于HMOS的新糖缀合物与诺如病毒衣壳的结合取决于特定的病毒株特征，这意味着HMOS及其多价衍生物是预防诺如病毒的潜在抗粘附剂。
