A rapid and sensitive assay for neuraminidase using peanut lectin hemagglutination: application to Vibrio cholera and Trypanosoma cruzi.
A neuraminidase assay based on peanut lectin agglutination is described. Human red blood cells are used both as substrate for the enzyme and as probe for the lectin. The validity of the method is ascertained by measuring the enzyme and lectin activities on erythrocytes whose outer membrane sialic acid was labeled with tritium after oxidation with sodium periodate followed by reduction with sodium borotritiide. The neuraminidases of Trypanosoma cruzi and Vibrio cholera are used as examples; in both cases, a linear relationship is observed between the degree of erythrocyte desialylation and the peanut hemagglutination titer. For the hemagglutination assay, lectin in homogeneous form as well as in crude peanut extracts may be used, and free sialic acid need not be separated from substrate-bound sialic acid. The hemagglutinating activity of peanut lectin is not affected by pre-treatment of the erythrocytes with various proteases. The method is particularly useful in the neuraminidase analysis of multiple samples, such as in gel filtration chromatography and for screening of hybridoma antibodies against neuraminidase.
D002850:Chromatography, Gel; D006386:Hemagglutination Tests / Q000379:methods*; D006801:Humans; D037102:Lectins; D009439:Neuraminidase / Q000032:analysis* / Q000378:metabolism; D019887:Peanut Agglutinin; D014349:Trypanosoma cruzi / Q000201:enzymology*; D014734:Vibrio cholerae / Q000201:enzymology*
D037102:Lectins; D019887:Peanut Agglutinin; D009439:Neuraminidase