A Boronic Acid Assay for the Detection of Mucin‐1 Glycoprotein from Cancer Cells. (19th July 2017)

• Record Type: Journal Article
• Authors: Zhang, Xiaoyu; Zhang, Shiqiang; Baek, Seung Joon; Best, Michael D.
• Is Part Of: Chembiochem. Volume 18:Number 16(2017)
• Page Start: 1578
• Page End: 1582
• Publication Date: 2017-07-19
• Subjects: boronic acid -- glycoproteins -- host–guest systems -- microarrays -- sensors; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201700288
• Languages: English
• ISSNs: 1439-4227

Abstract

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 acid1 onto streptavidin plates generated a multivalent surface for glycoprotein recruitment and detection. We first validated the binding properties of1 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. Abstract : 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.