Glycan (carbohydrate microarrays), carrying tens or hundreds of different sugars that are immobilized on solid surfaces in a spatially discrete pattern, possess a wide variety of potential applications in glycomics including rapid determination of the binding profile of carbohydrate-binding proteins, detection of specific antibodies for the diagnosis of diseases, characterization of carbohydrate-cell recognition events and the high-throughput screening of inhibitors to prevent carbohydrate-protein interactions. However, platforms that would enable construction of high quality carbohydrate microarrays and sensitive and rapid determination and characterization of carbohydrate-protein interactions have not kept pace with, for example, equivalent technology in genomic and proteomic research. To address this need, we have developed a unique novel 3-dimensional platform which we have demonstrated successfully for fabrication of high sensitive carbohydrate microarray in Phase I research;this platform could potentially allow us to screen thousands of different carbohydrate probes for binding to proteins including receptors and antibodies. In the Phase II project, we propose to build on our feasibility work to develop advanced carbohydrate microarray platform for high throughput glycan analysis. Specifically, we propose to (1) optimize a 3D glass-substrate microarray platform that will function with the existing fluorescent-based glycan analysis methods;(2) systematically compare of the effect of platform surface chemistry to glycan functions and epitope-displays;(3) develop epitope-defined high avidity N-glycan microarray for prostate cancer glycan-based biomarkers discovery, and (4) to develop 3D SPR substrate platform and a label-free surface plasma resonance (SPR) carbohydrate microarray system. The Phase II project results and deliverables will support the Phase II commercialization goals for implementing a Supercarbohydrate glass substrate for carbohydrate microarray fabrication that the fluorescent detection can be applied for the glycan analysis, and a Gold SPR platform substrate for carbohydrate microarray fabrication that the label-free detection can be applied for the glycan analysis. Follow-on products will include well-defined carbohydrate microarray for tumor biomarkers, and a label-free SPR imaging system for high throughput glycan analysis. The value of the platform substrates and the microarray system will bring to both the primary and secondary markets should attract large corporations as partners to support commercial development, marketing, and distribution.
Interactions between carbohydrates and proteins define a wide range of biochemical mechanisms in the human body. At present there is no commercially viable tool for rapid screening and investigation of these carbohydrate interactions. The proposed work will develop and demonstrate a microarray tool to allow researchers to probe carbohydrate biochemistry to elucidate disease and drug mechanisms and identify cancer biomarkers.
