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Chembiochem. 2017 Aug 17;18(16):1578-1582. doi: 10.1002/cbic.201700288. Epub 2017 Jul 19.

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

Author information

1
Department of Chemistry, University of Tennessee, 1420 Circle Drive, Knoxville, TN, 37996, USA.
2
Department of Biomedical and Diagnostic Sciences, College of Veterinary Medicine, University of Tennessee, 2407 River Drive, Knoxville, TN, 37996, USA.
3
Laboratory of Signal Transduction, College of Veterinary Medicine and, Research Institute for Veterinary Science, Seoul National University, Seoul, 08826, Republic of Korea.

Abstract

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.

KEYWORDS:

boronic acid; glycoproteins; host-guest systems; microarrays; sensors

PMID:
28574628
DOI:
10.1002/cbic.201700288
[Indexed for MEDLINE]

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