Recent research is uncovering unexpected ways that glycans contribute to biology, as well as new strategies for combatting diseases using approaches involving glycans. To make full use of the opportunities that are opening, we need methods that will enable biologists and clinicians who are not experts in glycobiology to conduct research in this field. The current methods require a high degree of expertise or specialized equipment. The methods also typically do not provide information relevant to disease-oriented research, such as the location, nature, and precise levels of specific glycans in clinical specimens. The goal of this project is to develop a method that will help meet this need. The method, which we call On-Chip Glycan Modification and Probing (On-Chip GMAP), will be accessible to non-specialists, usable with low sample volumes, and effective in providing precise measurements over many samples. We previously demonstrated micro-scale capture and affinity-based probing of glycans and glycoproteins from biological samples, as well as algorithms and software for interpreting the data. Here we will add enzymatic alterations of the captured glycans, followed by lectin/antibody probing of the altered glycans and automated interpretation of the data. The protocols are straightforward; reagents can be supplied in kit form; costs can be kept low because of the miniaturized scale; and the interpretation can be made readily understandable through the software. The information will be complementary to that derived from conventional glycan analyses using mass spectrometry and chromatography. Mass spectrometry and chromatography provide compositions and candidate structures for a limited number of samples, and the new method will provide motif information in a quantitative way over many samples. Therefore, On-Chip GMAP could be an important component of practical and comprehensive glycan analysis strategies, particularly for disease research. In addition, it could broaden the use of glycan analysis tools to researchers who are not experts in glycobiology, potentially facilitating new discoveries and applications.
Currently, a relatively small group of researchers pursues studies involving glycobiology, owing to the expertise and specialized tools required for the research. To enable more rapid and broader advances in glycobiology and its applications, this project aims to provide a practical method for non-specialists. The new method has the potential to be well suited to disease-oriented research and to become a critical component of comprehensive, practical glycan-analysis strategies.
| Klamer, Zachary; Staal, Ben; Prudden, Anthony R et al. (2017) Mining High-Complexity Motifs in Glycans: A New Language To Uncover the Fine Specificities of Lectins and Glycosidases. Anal Chem 89:12342-12350 |
| Barnett, Daniel; Liu, Ying; Partyka, Katie et al. (2017) The CA19-9 and Sialyl-TRA Antigens Define Separate Subpopulations of Pancreatic Cancer Cells. Sci Rep 7:4020 |
| Reatini, Bryan S; Ensink, Elliot; Liau, Brian et al. (2016) Characterizing Protein Glycosylation through On-Chip Glycan Modification and Probing. Anal Chem 88:11584-11592 |
