title: An enzyme-linked lectin assay for sialidase.
abstract: A procedure for the detection of low activities of sialidase (= neuraminidase) is described. Natural substrates for sialidase (human erythrocytes, fetuin or gangliosides) were coated onto the wells of microplates and incubated at 37 degrees C with the enzyme. Sialidase-induced desialylation of these natural substrates unmasks saccharides that are specifically recognized by the peanut agglutinin lectin (PNA). The use of a peroxidase-conjugated PNA (Po-PNA) allowed the binding of the lectin to the desialylated substrate to be quantified. The amount of bound Po-PNA correlated directly with the amount of sialic acid released from the substrate, and therefore with the sialidase activity. With this method, it was possible to detect sialidase activity associated with bacteria, myxoviruses and cells from higher organisms. This method may have important clinical implications as the use of ELISA allows automation and concurrent analysis of numerous samples.
authors.affiliation: INSERM U 139, Hopital H. Mondor, Créteil, France.
authors.forename: C R
authors.initials: CR
authors.lastname: Lambré
authors.forename: H
authors.initials: H
authors.lastname: Terzidis
authors.forename: A
authors.initials: A
authors.lastname: Greffard
authors.forename: R G
authors.initials: RG
authors.lastname: Webster
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004912:Erythrocytes / Q000378:metabolism; D005732:Gangliosides / Q000378:metabolism; D006168:Guinea Pigs / Q000097:blood; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D037102:Lectins / Q000378:metabolism*; D008264:Macrophages / Q000201:enzymology; D009439:Neuraminidase / Q000378:metabolism*; D009975:Orthomyxoviridae / Q000201:enzymology; D019887:Peanut Agglutinin; D014734:Vibrio cholerae / Q000201:enzymology; D000509:alpha-Fetoproteins / Q000378:metabolism
chemical_list: D005732:Gangliosides; D037102:Lectins; D019887:Peanut Agglutinin; D000509:alpha-Fetoproteins; D009439:Neuraminidase