用于糖芯片构建的人乳寡糖分离制备及其序列结构研究

人乳寡糖是人乳主要活性功能成分之一，对婴儿健康成长发育非常重要，有抗病原菌粘附、抗炎、免疫调节等作用，已成为近年来食品科学、营养学等领域关注的焦点之一。由于来源和手段的限制，加之结构复杂，现有人乳寡糖研究多以混合物为材料，能用于高通量糖芯片功能筛选的结构明确的微量人乳寡糖单体组分分离制备研究较少，制约了在糖链-蛋白相互作用的分子水平上阐明人乳寡糖作用机制的研究。本项目拟在前期研究基础上，运用色谱、质谱及色质联用等技术，进一步开展异构体水平的基于吡唑啉酮类异二官能团试剂标记的人乳寡糖高效制备及序列结构鉴定研究，构建一定规模的结构明确的人乳寡糖库和质谱结构数据库，用于人乳寡糖芯片的构建。重点解决唾液酸化人乳寡糖异构体的高效制备与质谱分析鉴定及后续糖芯片构建的兼容性问题。本项目将为深入研究人乳寡糖结构与功能的关系，进一步阐明人乳寡糖作用机制奠定基础，为研发营养健康的婴幼儿配方食品提供理论依据。

Human milk oligosaccharides(HMOs) are one of the main biological active ingredients in human milk, which play significant roles in breast-fed neonates’ development and growth. Numerous studies have shown that HMOs exert broad beneficial effects, including promoting the growth of gut beneficial organisms, prevention against viral infections and pathogen adhesion, and modulating the immune responses. Most results of the current studies obtained are based on HMOs mixture due to the lack of diversity HMO monomers, which limit the research on their structure - function relationships. This program will develop an integrative technology for HMO labeling (using pyrazolone-type heterobifunctional chromogenic reagents as the derivatization substrates), separation and preparation of HMO monomers, and construction of microarray for functional analysis. The prepared HMO monomers will be analyzed by LC-MS/MSn and MALDI-TOF-MS to elucidate their molecular weights, structural characterization, and sequences. The HMOs structural-activity relationship is expected to be explored as well. This program will be helpful to further illustrate the mechanism of HMOs activity and provide a theoretical basis for the basic research and development of healthy infant formula food.