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Title 

Semi-quantitative measurement of a specific glycoform using a DNA-tagged antibody and lectin affinity chromatography for glyco-biomarker development

Authors 

Lee Ju HeeCho chang-heeKim Sun HeeJeong Gu KangJeong Heon KoYong Sam Kim

Publisher 

American Society for Biochemistry and Molecular Biology

Issue Date 

2015

Citation 

Molecular & Cellular Proteomics, vol. 14, no. 3, pp. 782-795

Abstract 

Aberrant glycosylation-targeted disease biomarker development is based on cumulative evidence that certain glycoforms are mass-produced in a disease-specific manner. However, the development process has been hampered by the absence of an efficient validation method based on a sensitive and multiplexed platform. In particular, ELISA-based analytical tools are not adequate for this purpose, mainly because of the presence of a pair of N-glycans of IgG-type antibodies. To overcome the associated hurdles in this study, antibodies were tagged with oligonucleotides with T7 promoter and then allowed to form a complex with corresponding antigens. An antibody-bound specific glycoform was isolated by lectin chromatography and quantitatively measured on a DNA microarray chip following production of fluorescent RNA by T7-trascription. This tool ensured measurement of targeted glycoforms of multiple biomarkers with high sensitivity and multiplexity. This analytical method was applied to an in vitro diagnostic multivariate index assay where a panel of hepatocellular carcinoma (HCC) biomarkers comprising alpha-fetoprotein, hemopexin, and alpha-2-macroglobulin (A2M) was examined in terms of the serum level and their fuco-fractions. The results indicated that the tests using the multiplexed fuco-biomarkers provided improved discriminatory power between non-hepatocellular carcinoma and hepatocellular carcinoma subjects compared with the alpha-fetoprotein level or fuco-alphafetoprotein test alone. The developed method is expected to facilitate the validation of disease-specific glycan biomarker candidates.

ISSN 

1535-9476

Link 

http://dx.doi.org/10.1074/mcp.O114.043117

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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