SBIR PHASE I. TOPIC321:CHEMICALLY DEFINED GLYCAN LIBRARIES FOR REFERENCE STANDARD

鞘糖脂是人体细胞膜的重要组成部分。300多 已经表征了鞘糖脂聚糖。聚糖直接参与分子 识别事件。正在开发许多与癌症相关的鞘糖脂聚糖， 癌症疫苗大多数鞘糖脂聚糖不可商购。有限的成本 商业上可获得的数量非常高。我们开发了高效的一锅法 多酶（OPME）方法，用于高产率和成本效益的生产复合聚糖。的 OPME方法将用于产生60种结构不同的鞘糖脂聚糖库 包括含有不同天然唾液酸形式的那些。三个具体目标是 合成1）神经节系列; 2）乳糖和新乳糖系列; 3）球蛋白和异球蛋白系列 使用我们建立的高效OPME策略，分析鞘糖脂聚糖。每60个 化合物将在GLP条件下以10-50 mg规模合成，纯化至>98%纯度， 通过核磁共振光谱（NMR）和高分辨率质谱（MS）表征， 光谱法（HRMS）。对于合成的所有聚糖（纯度>98%），将每种50微克发送至 NIGMS设计的筛选中心，用于验证测试、聚糖微阵列和其他筛选 测定。这些聚糖是生物测定和生物医学的基本标准和宝贵的探针 应用.

Glycosphingolipids are essential components of human plasma membrane. More than 300 glycosphingolipid glycans have been characterized. The glycans are directly involved in molecular recognition event. A number of cancer-associated glycosphingolipid glycans are being developed as cancer vaccines. Most glycosphingolipid glycans are not commercially available. The costs for a limited number of commercially available ones are extremely high. We have developed highly effective one-pot multienzyme (OPME) approaches for high-yield and cost-effective production of complex glycans. The OPME approaches will be used in generating a library of 60 structurally diverse glycosphingolipid glycans including those contain different naturally occurring sialic acid forms. Three specific aims are to synthesize 1) ganglio-series; 2) lacto- and neolacto-series; and 3) globo- and isoglobo-series glycosphingolipid glycans using our established high efficient OPME strategies. Each of the 60 compounds will be synthesized under GLP conditions in 10-50 mg scale, purified to >98% purity, and characterized by nuclear magnetic resonance spectroscopy (NMR) and high-resolution mass spectrometry (HRMS). For all glycans synthesized (>98% purityL 50 micrograms of each will be sent to an NIGMS-designed screening center for validation testing, glycan microarray and other screening assays. These glycans are essential standards and invaluable probes for bioassays and biomedical applications.