title: Identification of IgM as a contaminant in lectin-FLISA assays for HCC detection.
abstract: 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.
authors.affiliation: Drexel University College of Medicine, Department of Microbiology and Immunology, Room 18307 New College Building, 245 N. 15th Street, Philadelphia, PA, 19102, USA.
authors.forename: Mengjun
authors.initials: M
authors.lastname: Wang
authors.affiliation: Drexel University College of Medicine, Department of Microbiology and Immunology, Room 18307 New College Building, 245 N. 15th Street, Philadelphia, PA, 19102, USA.
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Comunale
authors.affiliation: Graduate School of Biomedical Sciences and Professional Studies, Drexel University College of Medicine, Microbiology and Immunology Graduate Program, 2900 Queen Lane, Philadelphia, PA, 19129, USA.
authors.forename: Harmin
authors.initials: H
authors.lastname: Herrera
authors.affiliation: Drexel University College of Medicine, Department of Microbiology and Immunology, Room 18307 New College Building, 245 N. 15th Street, Philadelphia, PA, 19102, USA.
authors.forename: Lucy
authors.initials: L
authors.lastname: Betesh
authors.affiliation: Division of Gastroenterology & Hepatology, University of California, San Diego 200 West Arbor Drive, San Diego, CA 92103-8413, USA.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kono
authors.affiliation: Drexel University College of Medicine, Department of Microbiology and Immunology, Room 18307 New College Building, 245 N. 15th Street, Philadelphia, PA, 19102, USA. Electronic address: anand.mehta@drexelmed.edu.
authors.forename: Anand
authors.initials: A
authors.lastname: Mehta
mesh_terms: D014408:Biomarkers, Tumor / Q000097:blood; D006528:Carcinoma, Hepatocellular / Q000097:blood*; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000097:blood; D007075:Immunoglobulin M / Q000032:analysis / Q000097:blood*; D007163:Immunosorbent Techniques; D037102:Lectins / Q000032:analysis*; D008113:Liver Neoplasms / Q000097:blood*; D000509:alpha-Fetoproteins / Q000032:analysis*
chemical_list: D014408:Biomarkers, Tumor; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D037102:Lectins; D000509:alpha-Fetoproteins
keywords: Biomarker; Glycosylation; Hepatocellular carcinoma; Lectin; Liver cancer