Novel mouse monoclonal antibodies specifically recognize Aspergillus fumigatus galactomannan

Andrey L. Matveev, Vadim B. Krylov, Ljudmila A. Emelyanova, Arsenii S. Solovev, Yana A. Khlusevich, Ivan K. Baykov, Thierry Fontaine, Jean Paul Latgé, Nina V. Tikunova, Nikolay E. Nifantiev

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

Abstract

A panel of specific monoclonal antibodies (mAbs) against synthetic pentasaccharide β-D-Galf-(1!5)-[β-D-Galf-(1!5)]3-α-D-Manp, structurally related to Aspergillus fumigatus galactomannan, was generated using mice immunized with synthetic pentasaccharide-BSA conjugate and by hybridoma technology. Two selected mAbs, 7B8 and 8G4, could bind with the initial pentasaccharide with affinity constants of approximately 5.3 nM and 6.4 nM, respectively, based on surface plasmon resonance-based biosensor assay. The glycoarray, built from a series of synthetic oligosaccharide derivatives representing different galactomannan fragments, demonstrated that mAb 8G4 could effectively recognize the parental pentasaccharide while mAb 7B8 recognizes its constituting trisaccharide parts. Immunofluorescence studies showed that both 7B8 and 8G4 could stain A. fumigatus cells in culture efficiently, but not the mutant strain lacking galactomannan. In addition, confocal microscopy demonstrated that Candida albicans, Bifidobacterium longum, Lactobacillus plantarum, and numerous gram-positive and gram-negative bacteria were not labeled by mAbs 7B8 and 8G4. The generated mAbs can be considered promising for the development of a new specific enzyme-linked assay for detection of A. fumigatus, which is highly demanded for medical and environmental controls.

Original languageEnglish
Article number0193938
Pages (from-to)e0193938
Number of pages15
JournalPLoS ONE
Volume13
Issue number3
DOIs
Publication statusPublished - 8 Mar 2018

Keywords

  • Animals
  • Antibodies, Fungal/immunology
  • Antibodies, Monoclonal/immunology
  • Antibody Specificity
  • Antigens, Fungal/immunology
  • Aspergillus fumigatus/immunology
  • Biotinylation
  • Carbohydrate Sequence
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Fluorescent Antibody Technique, Direct
  • Hybridomas/immunology
  • Mannans/chemical synthesis
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Confocal
  • Oligosaccharides/chemical synthesis
  • INFECTIONS
  • PERFORMANCE
  • TESTING SERUM
  • INVASIVE ASPERGILLOSIS
  • CROSS-REACTIVITY
  • ENZYME-IMMUNOASSAY
  • LINKED-IMMUNOSORBENT-ASSAY
  • IDENTIFICATION
  • INTO-FURANOSIDE REARRANGEMENT
  • PCR

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