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Biomarker analysis of fucosylated kininogen through depletion of lectin reactive heterophilic antibodies in hepatocellular carcinoma
Journal article   Open access   Peer reviewed

Biomarker analysis of fucosylated kininogen through depletion of lectin reactive heterophilic antibodies in hepatocellular carcinoma

Mengjun Wang, Jiabin Shen, Harmin Herrera, Amit Singal, Charles Swindell, Lu Renquan and Anand Mehta
Journal of immunological methods, v 462
Nov 2018
PMID: 30144410
url
https://europepmc.org/articles/pmc6784319View
Accepted (AM)Open Access (License Unspecified) Open

Abstract

Biomarker Glycosylation Hepatocellular carcinoma Lectin, liver cance
Hepatocellular carcinoma (HCC) accounts for >700,000 deaths worldwide, largely related to poor rates of diagnosis. Our previous work identified glycoproteins with increased levels of fucosylation in HCC. Plate-based assays to measure this change were compromised by increased levels of heterophilic antibodies with glycan lacking terminal galactose residues, which allowed for increased binding to the lectins used in these assays. To address this issue, we developed a multi-step protein A/G incubation and filtration method to remove the contaminating signal. However, this method was time consuming and expensive so alternative methods were desired. Herein, we describe a simple method relying on PEG precipitation that allows for the removal of IgG and IgM but retention of glycoproteins of interest. This method was tested on three sample sets, two internal and one external. PEG depletion of heterophilic IgG and IgM reduced in the coefficient of variation as observed with the protein A/G filtration method from 26.82% to 7.50% and allowed for the measurement of fucosylated protein. This method allowed for the measurement of fucosylated kininogen, which could serve as a biomarker of HCC. In conclusion, a new and simple method for the depletion of heterophilic IgG and IgM was developed and allowed for the analysis of fucosylated kininogen in patients with liver disease. •Developed a method for the depletion of lectin reactive heterophilic antibodies•This method allows for the analysis of fucosylated proteins in serum.•This method reduced the coefficient of variation of the assay.•Specific fucosylated proteins can act as biomarkers of hepatocellular carcinoma.•Biomarker containing a fucosylated protein has excellent discriminatory.

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Collaboration types
Domestic collaboration
International collaboration
Web of Science research areas
Biochemical Research Methods
Immunology
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