Rat monoclonal antibodies against Aspergillus galactomannan.
Monoclonal antibodies (MAbs) against Aspergillus fumigatus galactomannan were produced in rats. Seven of them, EB-A1 through EB-A7, were characterized in more detail. They were all immunoglobulin M antibodies, reacting in an indirect enzyme-linked immunosorbent assay with purified A. fumigatus galactomannan, with avidity constants of between 2 x 10(9) and 5 x 10(9)/M. Enzyme-linked immunosorbent assay inhibition experiments with modified galactomannan and synthetic oligomers of beta (1----5)galactofuranose demonstrated that the MAbs bound to an epitope located on the beta(1----5)galactofuranose-containing side chains of the galactomannan molecule. An identical or similar epitope also seemed to be present in other fungi. Immunofluorescence and immunoelectron microscopy experiments with EB-A2 revealed the presence of the antigen in the fungal wall and inside the cell. Immunoblotting experiments demonstrated that the epitope recognized by the MAbs was a common oligosaccharide moiety of a wide range of intracellular and extracellular glycoproteins in A. fumigatus. The characteristics of the MAbs justify their use in the diagnosis of invasive aspergillosis by antigen detection.
Stynen
D
D
Sanofi Diagnostics Pasteur, Genk, Belgium.
Sarfati
J
J
Goris
A
A
Prévost
M C
MC
Lesourd
M
M
Kamphuis
H
H
Darras
V
V
Latgé
J P
JP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000946:Antigens, Fungal / Q000032:analysis; D001230:Aspergillus / Q000276:immunology*; D003429:Cross Reactions; D000939:Epitopes / Q000032:analysis; D005455:Fluorescent Antibody Technique; D005690:Galactose / Q000031:analogs & derivatives; D015151:Immunoblotting; D008351:Mannans / Q000276:immunology*; D051381:Rats
D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D000939:Epitopes; D008351:Mannans; C012990:galactomannan; D005690:Galactose