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So You Discovered a Potential Glycan-Based Biomarker; Now What? We Developed a High-Throughput Method for Quantitative Clinical Glycan Biomarker Validation
Shipman, Joshua T. Nguyen, Hanna T. Desaire, Heather
Shipman, Joshua T.
Nguyen, Hanna T.
Desaire, Heather
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Abstract
Glycomic-based approaches to discover potential biomarkers have shown great promise in their ability to distinguish between healthy and diseased individuals; these methods can identify when aberrant glycosylation is significant, but they cannot practically be adapted into widely implemented diagnostic assays because they are too complex, expensive, and low-throughput. We have developed a new strategy that addresses challenges associated with sample preparation, sample throughput, instrumentation needs, and data analysis to transfer the valuable knowledge provided by protein glycosylation into a clinical environment. Notably, the detection limits of the assay are in the single-digit picomole range. Proof of principle is demonstrated by quantifying the changes in the sialic acid content in fetuin. As the sialic acid content in proteins varies in a number of disease states, this example demonstrates the utility of the method for biomarker analysis. Furthermore, the developed method can be adapted to other biologically important saccharides, affording a broad array of quantitative glycomic analyses that are accessible in a high-throughput, plate-reader format. These studies enable glycomic-based biomarker discovery efforts to transition through the difficult landscape of developing a potential biomarker into a clinical assay.
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Date
2020-03-18
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American Chemical Society
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Keywords
Assays, Carbohydrates, Chemical biology, Extraction, Fluorescence
Citation
Joshua T. Shipman, Hanna T. Nguyen, and Heather Desaire. ACS Omega 2020 5 (12), 6270-6276. DOI: 10.1021/acsomega.9b03334