ChemBioChem
ChemBioChem
5 years ago

A Boronic Acid Assay for the Detection of Mucin-1 Glycoprotein from Cancer Cells

A Boronic Acid Assay for the Detection of Mucin-1 Glycoprotein from Cancer Cells
Seung Joon Baek, Shiqiang Zhang, Michael D. Best, Xiaoyu Zhang
Cell surface glycoproteins are commonly aberrant in disease and act as biomarkers that facilitate diagnostics. Mucin-1 (MUC1) is a prominent example, exhibiting truncated glycosylation in cancer. We present herein a boronic acid microplate assay for sensitive and high-throughput detection of such glycoproteins. The immobilization of biotin–boronic acid 1 onto streptavidin plates generated a multivalent surface for glycoprotein recruitment and detection. We first validated the binding properties of 1 in solution through titrations with alizarin dye. Next, the microplate assay was explored through horseradish peroxidase (HRP) analysis as a proof-of-concept glycoprotein with chemiluminescence detection. Finally, this platform was applied for the detection of MUC1 directly from MCF-7 human breast cancer cell lysates by using an HRP-tagged antibody that targets the cancerous form of this glycoprotein. Sensitive, dose-dependent detection of MUC1 was observed, showcasing the efficacy of this platform for detecting disease-associated glycoproteins. Disease detection: Glycoproteins such as mucin-1 commonly exhibit aberrant glycosylation patterns in diseased cells, thereby providing a marker for diagnosis. Immobilizing boronic acid–biotin sensors onto streptavidin microplates resulted in a multivalent surface for sensitive and high-throughput detection of glycoproteins by using chemiluminescence signal transduction, which was applied to directly detect mucin-1 in cancer cell lysates.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/cbic.201700288

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