Monoclonal Antibody AP3 Binds Galactomannan Antigens Displayed by the Pathogens Aspergillus flavus, A. fumigatus, and A. parasiticus

Aspergillus fumigatus and A. flavus are the fungal pathogens responsible for most cases of invasive aspergillosis (IA). Early detection of the circulating antigen galactomannan (GM) in serum allows the prompt application of effective antifungal therapy, thus improving the survival rate of IA patients. However, the use of monoclonal antibodies (mAbs) for the diagnosis of IA is often associated with false positives due to cross-reaction with bacterial polysaccharides. More specific antibodies are therefore needed. Here we describe the characterization of the Aspergillus-specific mAb AP3 (IgG1κ), including the precise identification of its corresponding antigen. The antibody was generated using A. parasiticus cell wall fragments and was shown to bind several Aspergillus species. Immunofluorescence microscopy revealed that AP3 binds a cell wall antigen, but immunoprecipitation and enzyme-linked immunosorbent assays showed that the antigen is also secreted into the culture medium. The inability of AP3 to bind the A. fumigatus galactofuranose (Galf )-deficient mutant ΔglfA confirmed that Galf residues are part of the epitope. Several lines of evidence strongly indicated that AP3 recognizes the Galf residues of O-linked glycans on Aspergillus proteins. Glycoarray analysis revealed that AP3 recognizes oligo-[β-D-Galf-1,5] sequences containing four or more residues with longer chains more efficiently. We also showed that AP3 captures GM in serum, suggesting it may be useful as a diagnostic tool for patients with IA.

• Publication year

  2019

• Venue

  Frontiers in Cellular and Infection Microbiology

• Publication date

  2019-07-16

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.3389/fcimb.2019.00234PMID 31380292PMCID 6646516
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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• Convergent synthesis of isomeric heterosaccharides related to the fragments of galactomannan from Aspergillus fumigatus.

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• A monoclonal antibody against glycoproteins of Aspergillus fumigatus shows anti-adhesive potential.

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• Immunodetection of fungal and oomycete pathogens: Established and emerging threats to human health, animal welfare and global food security

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• Significant structural change in both O- and N-linked carbohydrate moieties of the antigenic galactomannan from Aspergillus fumigatus grown under different culture conditions.

  2015cited by this paper
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• Aflatoxins and safe storage

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• Phylogeny, identification and nomenclature of the genus Aspergillus

  2014cited by this paper
• Transcriptional activation of heat shock protein 90 mediated via a proximal promoter region as trigger of caspofungin resistance in Aspergillus fumigatus.

  2014cited by this paper
• Heat shock protein 90 (Hsp90): A novel antifungal target against Aspergillus fumigatus

  2014cited by this paper
• Methods for Detection of Aflatoxins in Agricultural Food Crops

  2014cited by this paper
• Simultaneous treatment with tebuconazole and abscisic acid induces drought and salinity stress tolerance in Arabidopsis thaliana by maintaining key plastid protein levels.

  2013cited by this paper
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  2013cited by this paper
• gfsA encodes a novel galactofuranosyltransferase involved in biosynthesis of galactofuranose antigen of O‐glycan in Aspergillus nidulans and Aspergillus fumigatus

  2013cited by this paper
• Galactofuranose in eukaryotes: aspects of biosynthesis and functional impact.

  2012cited by this paper
• Heat Shock Protein 90 Is Required for Conidiation and Cell Wall Integrity in Aspergillus fumigatus

  2012cited by this paper
• Studies on galactofuranose-containing glycostructures of the pathogenic mold Aspergillus fumigatus.

  2011cited by this paper
• Cross-Reactivity of Fusarium spp. in the Aspergillus Galactomannan Enzyme-Linked Immunosorbent Assay

  2011cited by this paper
• One-Step Protein Purification: Use of a Novel Epitope Tag for Highly Efficient Detection and Purific

  2011cited by this paper
• Proteome profiling and functional classification of intracellular proteins from conidia of the human-pathogenic mold Aspergillus fumigatus.

  2010cited by this paper
• Detection of invasive aspergillosis.

  2010influential reference
• Cultivation characteristics and gene expression profiles of Aspergillus oryzae by membrane-surface liquid culture, shaking-flask culture, and agar-plate culture.

  2010cited by this paper
• Protection by Anti-β-Glucan Antibodies Is Associated with Restricted β-1,3 Glucan Binding Specificity and Inhibition of Fungal Growth and Adherence

  2009cited by this paper
• Essentials of glycobiology

  2009cited by this paper
• Aspergillus flavus: an emerging non‐fumigatus Aspergillus species of significance

  2009cited by this paper
• Galactofuranose containing molecules in Aspergillus fumigatus.

  2009cited by this paper
• The Fusarium oxysporum cell wall proteome under adhesion‐inducing conditions

  2009influential reference
• Effects of glycosylation on the stability of protein pharmaceuticals.

  2009cited by this paper
• Scientific Advances with Aspergillus Species that Are Used for Food and Biotech Applications.

  2008cited by this paper
• Contribution of Galactofuranose to the Virulence of the Opportunistic Pathogen Aspergillus fumigatus

  2008cited by this paper
• Development of an Immunochromatographic Lateral-Flow Device for Rapid Serodiagnosis of Invasive Aspergillosis

  2008cited by this paper
• False-Positive Results by the Platelia Aspergillus Galactomannan Antigen Test for Patients Treated with Amoxicillin-Clavulanate

  2008cited by this paper
• Cell wall fractionation for yeast and fungal proteomics.

  2008cited by this paper
• Treatment of aspergillosis: clinical practice guidelines of the Infectious Diseases Society of America.

  2008cited by this paper
• Well-Characterized Monoclonal Antibodies against Cell Wall Antigen of Aspergillus Species Improve Immunoassay Specificity and Sensitivity

  2007cited by this paper
• Aspergillus flavus: human pathogen, allergen and mycotoxin producer.

  2007cited by this paper
• Occurrence and Kinetics of False-Positive Aspergillus Galactomannan Test Results following Treatment with β-Lactam Antibiotics in Patients with Hematological Disorders

  2006cited by this paper
• Diagnosis of invasive aspergillosis using a galactomannan assay: a meta-analysis.

  2006cited by this paper
• Aspergillus Infections in Transplant Recipients

  2005cited by this paper
• Cryptococcus neoformans Galactoxylomannan Contains an Epitope(s) That Is Cross-Reactive with Aspergillus Galactomannan

  2005cited by this paper
• Bifidobacterial Lipoglycan as a New Cause for False-Positive Platelia Aspergillus Enzyme-Linked Immunosorbent Assay Reactivity

  2005cited by this paper
• False-positive galactomannan platelia Aspergillus test results for patients receiving piperacillin-tazobactam.

  2004cited by this paper
• AFMP2 Encodes a Novel Immunogenic Protein of the Antigenic Mannoprotein Superfamily in Aspergillus fumigatus

  2004cited by this paper
• Catalases of Aspergillus fumigatus

  2003cited by this paper
• AFLMP1 Encodes an Antigenic Cell Wall Protein in Aspergillus flavus

  2003cited by this paper
• The Costs of Mycotoxin Management to the USA: Management of Aflatoxins in the United States

  2003cited by this paper
• The 5-O-/3-D-Galactofuranosyl-containing Glycopeptide from Penicillium charlesii

  2002cited by this paper
• Detection of Antibodies Specific to an Antigenic Cell Wall Galactomannoprotein for Serodiagnosis of Aspergillus fumigatus Aspergillosis

  2002cited by this paper
• Aspergillus fumigatus cell wall: composition and biosynthesis.

  2001cited by this paper
• Proteome analysis of Aspergillus fumigatus identifies glycosylphosphatidylinositol‐anchored proteins associated to the cell wall biosynthesis

  2001cited by this paper
• The impact of culture isolation of Aspergillus species: a hospital-based survey of aspergillosis.

  2001cited by this paper
• Fungal allergens and peptide epitopes.

  2000cited by this paper
• Chemical and immunological characterization of the extracellular galactomannan of Aspergillus fumigatus

  1994cited by this paper
• Rat monoclonal antibodies against Aspergillus galactomannan

  1992influential reference
• Purification and characterisation of an extracellular serine proteinase from Aspergillus fumigatus and its detection in tissue.

  1990influential reference
• Production and characterization of monoclonal antibodies to cell wall antigens of Aspergillus fumigatus

  1990cited by this paper
• The 5-O-beta-D-galactofuranosyl-containing glycopeptide from Penicillium charlesii. Carbon 13 nuclear magnetic resonance studies.

  1979cited by this paper
• Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane.

  1971influential reference

• Recent developments in Aspergillus fumigatus research: diversity, drugs, and disease

  2025cites this paper
• Current trends in organic chemistry: contribution of the N. D. Zelinsky Institute of Organic Chemistry of the Russian Academy of Sciences

  2024cites this paper
• Galf-Specific Neolectins: Towards Promising Diagnostic Tools

  2024cites this paper
• Time-resolved fluoroimmunoassay for Aspergillus detection based on anti-galactomannan monoclonal antibody from stable cell line.

  2024cites this paper
• Synthesis of branched heterooligosaccharides related to Aspergillus galactomannan containing short Galf side chains.

  2024cites this paper
• Identification of a new DC-SIGN binding pentamannoside epitope within the complex structure of Candida albicans mannan

  2023cites this paper
• Anti-glycan monoclonal antibodies: Basic research and clinical applications.

  2023cites this paper
• Nano-biosensor based on the combined use of the dynamic and static light scattering for Aspergillus galactomannan analysis

  2022cites this paper
• Surfactant protein D inhibits growth, alters cell surface polysaccharide exposure and immune activation potential of Aspergillus fumigatus

  2022cites this paper
• Two Monoclonal Antibodies That Specifically Recognize Aspergillus Cell Wall Antigens and Can Detect Circulating Antigens in Infected Mice

  2021influential citation
• Anti-glycan antibodies: roles in human disease.

  2021cites this paper
• Selection and characterization of two monoclonal antibodies specific for the Aspergillus flavus major antigenic cell wall protein Aflmp1.

  2021cites this paper
• Reinvestigation of Carbohydrate Specificity of EBCA-1 Monoclonal Antibody Used for the Detection of Candida Mannan

  2021cites this paper
• Synthetic carbohydrate based anti-fungal vaccines.

  2020cites this paper
• Characterization of a novel yeast phase-specific antigen expressed during in vitro thermal phase transition of Talaromyces marneffei

  2020cites this paper
• Synthesis of Biotin-Tagged Chitosan Oligosaccharides and Assessment of Their Immunomodulatory Activity

  2020cites this paper
• Immunotherapeutic approaches for fungal infections.

  2020cites this paper
• Importance of Candida Antigenic Factors: Structure-Driven Immunomodulation Properties of Synthetically Prepared Mannooligosaccharides in RAW264.7 Macrophages

  2019cites this paper
• Potential of fluorescence polarization immunoassay for the detection of Aspergillus fumigatus galactomannan

  2019cites this paper