基于数字微流控平台的酶法合成糖链研究.

Glycans of glycoconjugates such as glycoproteins and glycolipids are very important for their proper functions. Investigation towards glycans has become the third milestone of exploring the mystery of life after proteins and nucleic acids. However, the structure-function relationships of glycans haven’t been fully established due to diversity and complexity of glycans, and challenge of access to pure glycans. Oligosaccharide synthesis is still the best choice at current stage. Thus, developing novel, efficient, economic and green technology for glycan synthesis will be of great theoretical significance and application value for investigation into the structure-function relationships of glycans, dyamic modification of glycosylated proteins and development of automated glycan synthesizers..In this project, in order to develop one novel technology for glycan synthesis based on DMF (digital microfluidics), the following studies will be carried out: DMF, magnetic beads, and enzymatic modular synthesis of glycans will be integrated to build an efficient and automated DMF-based platform for enzymatic modular synthesis of glycans; The application range of this platform for glycan synthesis will be further expanded by studying enzymatic modular synthesis of other oligosaccharides (including antigens such as 3’-sialyl Lewis and 3’-sialyl Lewis x, and human milk oligosaccharides like 2’-fucosyllactose, lacto-N-tetraose, and lacto-N-difucosylhexaose I) and polysaccharides such as hyaluronic acid. In the end, the DMF-based platform will be developed into one robust, highly efficient, economic, and automated technology for glycan synthesis.

糖链对糖复合物发挥正确功能非常关键，糖链研究已成为继蛋白质和核酸研究之后探索生命奥秘的第三个里程碑。由于糖链结构的多样性与复杂性以及纯糖链获得的挑战性，糖链的结构和功能之间还没有建立对应关系。合成仍是获得糖链的最佳选择，因此发展新型、高效、经济、绿色的糖链合成技术对糖链的结构-功能关系研究、糖链合成仪的研制等具有重要理论研究意义和实际应用价值。.本项目将以发展基于数字微流控（Digital Microfluidics，DMF）平台的酶催化合成糖链技术为目标，开展以下研究：首先以N-乙酰-alpha-2,6-唾液酸乳糖胺的酶催化合成为例，通过把DMF、磁珠以及酶催化模块合成糖链等技术集成，建立基于DMF的高效、自动酶催化合成糖链平台；通过研究其他功能寡糖及透明质酸（多糖）基于该平台的酶催化合成，进一步拓宽DMF糖链合成平台的应用范围，把该平台发展成为一种强大、高效、经济的自动糖链合成技术。

糖链对糖复合物发挥正确功能非常关键，糖链研究已成为继蛋白质和核酸研究之后探索生命奥秘的第三个里程碑。由于糖链结构的多样性与复杂性以及纯糖链获得的挑战性，糖链的结构和功能之间还没有建立对应关系。合成仍是获得糖链的最佳选择，因此发展新型、高效、经济、绿色的糖链合成技术对糖链的结构-功能关系研究、糖链合成仪的研制等具有重要理论研究意义和实际应用价值。. 本项目以发展基于数字微流控（Digital Microfluidics，DMF）平台的酶催化合成糖链技术为目标，开展了以下研究：建立了8个寡糖合成需要的酶催化模块；开发了基于超高频声微流控的生物素标记N-乙酰-alpha-2,6-唾液酸乳糖胺的酶催化合成技术；搭建了DMF、磁操控、泵输送等技术整合在的一起的糖链合成平台，并成功在该平台实现了多种寡糖、N-糖链及透明质酸（多糖）的合成，为全自动糖链合成仪的研制奠定了基础。