TY - JOUR

T1 - Novel mouse monoclonal antibodies specifically recognize Aspergillus fumigatus galactomannan

AU - Matveev, Andrey L.

AU - Krylov, Vadim B.

AU - Emelyanova, Ljudmila A.

AU - Solovev, Arsenii S.

AU - Khlusevich, Yana A.

AU - Baykov, Ivan K.

AU - Fontaine, Thierry

AU - Latgé, Jean Paul

AU - Tikunova, Nina V.

AU - Nifantiev, Nikolay E.

PY - 2018/3/8

Y1 - 2018/3/8

N2 - 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.

AB - 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.

KW - Animals

KW - Antibodies, Fungal/immunology

KW - Antibodies, Monoclonal/immunology

KW - Antibody Specificity

KW - Antigens, Fungal/immunology

KW - Aspergillus fumigatus/immunology

KW - Biotinylation

KW - Carbohydrate Sequence

KW - Enzyme-Linked Immunosorbent Assay

KW - Female

KW - Fluorescent Antibody Technique, Direct

KW - Hybridomas/immunology

KW - Mannans/chemical synthesis

KW - Mice

KW - Mice, Inbred BALB C

KW - Microscopy, Confocal

KW - Oligosaccharides/chemical synthesis

KW - INFECTIONS

KW - PERFORMANCE

KW - TESTING SERUM

KW - INVASIVE ASPERGILLOSIS

KW - CROSS-REACTIVITY

KW - ENZYME-IMMUNOASSAY

KW - LINKED-IMMUNOSORBENT-ASSAY

KW - IDENTIFICATION

KW - INTO-FURANOSIDE REARRANGEMENT

KW - PCR

UR - http://www.scopus.com/inward/record.url?scp=85043467382&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0193938

DO - 10.1371/journal.pone.0193938

M3 - Article

C2 - 29518144

AN - SCOPUS:85043467382

VL - 13

SP - e0193938

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 3

M1 - 0193938

ER -