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Identification of IgM as a contaminant in lectin-FLISA assays for HCC detection
Journal article   Open access   Peer reviewed

Identification of IgM as a contaminant in lectin-FLISA assays for HCC detection

Mengjun Wang, Mary Ann Comunale, Harmin Herrera, Lucy Betesh, Yuko Kono and Anand Mehta
Biochemical and biophysical research communications, v 476(3)
29 Jul 2016
DOI: https://doi.org/10.1016/j.bbrc.2016.05.027
PMID: 27181357
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://europepmc.org/articles/pmc5067009View
Accepted (AM) Open Open Access (License Unspecified)

Abstract

Biomarker Glycosylation Hepatocellular carcinoma Lectin Liver cancer
Liver disease, in the form of hepatocellular carcinoma (HCC) accounts for > 700,000 deaths worldwide. A major reason for this is late diagnosis of HCC. The currently used biomarker, serum alpha-fetoprotein (AFP) is elevated in 40–60% of those with HCC and other markers that can either compliment or replace AFP are desired. Our previous work has identified a number of proteins that contain altered glycans in HCC. Specifically, these altered glycans were increased levels of core and outer arm fucosylation. To determine the clinical usefulness of those identified glycoproteins, a plate based assay was developed that allowed for the detection of fucosylated glycoforms. While this method was applicable to a number of independent patient sets, it was unable to specifically detect fucosylated glycoforms in many patient samples. That is, some material was present in serum that led to non-specific signal in the lectin- fluorescence -linked immunosorbent assay (lectin-FLISA). To address this issue, a systematic process was undertaken to identify the material. This material was found to be increased levels of lectin reactive IgM. Removal of both IgG and IgM using a multi-step protein A/G incubation and filtration step removed the contaminating signal and allowed for the analysis of specific protein glycoforms. This assay was subsequently used on two sample sets, one that was shown previously to be unable to be tested via a lectin FLISA and in a larger independent sample set. The clinical usefulness of this assay in the early detection of HCC is discussed.

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Collaboration types
Domestic collaboration
Web of Science research areas
Biochemistry & Molecular Biology
Biophysics
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