Chromatically Orthogonal Photolabile Protecting Groups for the Parallel On-Chip Synthesis of High-Density Glycan Microarrays

用于并行片上合成高密度聚糖微阵列的色谱正交光不稳定保护基团

• 批准号: 10722250
• 负责人: Arthur H. Winter
• 金额: $ 14.86万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10722250
• 关键词: Acceleration; Antibodies; BODIPY; Bacteria; Benign; Binding; Binding Proteins; Biological Process; Carbohydrates; Collaborations; Color; Combinatorial Synthesis; Complex; DNA; DNA Sequence; Darkness; Development; Dyes; Epitope Mapping; Epitopes; Family; Fluorescence; Gene Chips; Genotype; Health; Human; Immune; In Situ; Individual; Label; Lectin; Libraries; Light; Matched Group; Methods; Monitor; Monosaccharides; Neoplasm Metastasis; Oligonucleotides; Outcomes Research; Photochemistry; Polymers; Polysaccharides; Reagent; Research; S phase; Signal Transduction; Source; Specificity; Structure; Structure-Activity Relationship; Surface; Technology; Viral; Virus; Work; absorption; chromophore; combinatorial; density; experience; experimental study; fabrication; gene synthesis; glycosylation; irradiation; manufacture; microchip; pathogen; screening; sugar; trafficking

Abstract. The objective of this research is to identify carbohydrate-compatible photolabile protecting groups and light sources to facilitate the parallel on-surface synthesis of high-density glycan microarrays—e.g. glycan chips—in a manner similar to the synthesis of gene chips. Instead of using monochromatic light and a single photolabile protecting group to spatially control the extension of a linear polymer (DNA) on the microchip, as in gene chip manufacture, irradiation with different wavelengths of light combined with wavelength-selective tem- porary protecting groups will allow for constructing complex, branched glycans on a microchip surface. I aim to: (1) identify carbohydrate-compatible photolabile protecting groups and optimal light sources to pair with each; and (2) identify a pair of wavelength-selective (`chromatically orthogonal') protecting groups and demonstrate their use in branched glycan synthesis, paving the way towards the parallel on-chip synthesis of high-density glycan microarrays. Wavelength-selective photochemistry will be achieved by separating the absorptions of pho- tolabile protecting groups to allow for selective excitation. Such high-density combinatorially-synthesized glycan chips are expected to permit rapid epitope mapping and screening of the selectivity of glycan binding partners. For example, exposing a dye-labeled lectin, antibody, or virus/bacteria/pathogen binding protein to the chip will allow its binding specificity for numerous glycan structures to be determined from a single experiment. The ability to synthesize high-density combinatorial libraries of carbohydrates will aid in resolving the structure-function relationships of carbohydrates, help to understand the target epitopes of glycan binding partners, and accelerate efforts to uncover the structure and function of the glycome.

摘要：本研究的目的是鉴定碳水化合物相容的光不稳定保护基团。 和光源以促进高密度聚糖微阵列的并行表面合成——例如聚糖 芯片——以类似于基因芯片合成的方式，而不是使用单色光和单一光。 光不稳定的保护基团可在空间上控制微芯片上线性聚合物 (DNA) 的延伸，如 基因芯片制造，不同波长的光照射结合波长选择温度 多孔保护基团将允许在微芯片表面构建复杂的支链聚糖，我的目标是： (1) 确定与碳水化合物相容的光不稳定保护基团以及与其配对的最佳光源； (2) 识别一对波长选择性（“色谱正交”）保护基团并证明 它们在支链聚糖合成中的应用，为高密度并行芯片合成铺平了道路 聚糖微阵列将通过分离光的吸收来实现。 不稳定的保护基团以允许选择性激发这种高密度组合合成的聚糖。 芯片有望实现快速表位作图和聚糖结合伴侣选择性筛选。 例如，将染料标记的凝集素、抗体或病毒/细菌/病原体结合蛋白暴露于芯片将 允许通过单个实验确定其对多种聚糖结构的结合特异性。 合成高密度碳水化合物组合库将有助于解决结构功能问题 碳水化合物的关系，有助于了解聚糖结合伙伴的目标表位，并加速 揭示糖组的结构和功能。