Quantitative determination of dietary lectin activities by enzyme-linked immunosorbent assay using specific glycoproteins immobilized on microtiter plates.
An immunoenzymatic method for the quantitative determination of dietary lectin activities employing immobilized glycoproteins was studied. Lectins from wheat germ (WGA), peanut (PNA), and jack bean (ConA) were added to microtiter plates coated with ovalbumin or asialofetuin and quantified by enzyme-linked immunosorbent assay (ELISA) with lectin-specific antibodies. ELISA responses for lectin activity were dose-dependent in the concentration range 30-1000 ng/mL for WGA and 80-1000 ng/mL for both PNA and ConA. Inhibition assays carried out with different saccharides confirmed that the binding of lectins to immobilized glycoproteins was specific. The proposed method is specific and sensitive, allowing the quantitative determination of lectin activities on raw samples by simple dilution of the extracts. Examples of application to wheat germ and roasted peanut extracts are reported.
Vincenzi
Simone
S
Dipartimento Scientifico e Tecnologico, Università di Verona, Strada Le Grazie 15, 37134, Verona, Italy.
Zoccatelli
Gianni
G
Perbellini
Fabio
F
Rizzi
Corrado
C
Chignola
Roberto
R
Curioni
Andrea
A
Peruffo
Angelo D B
AD
D000906:Antibodies / Q000276:immunology; D003208:Concanavalin A / Q000032:analysis / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006023:Glycoproteins / Q000378:metabolism*; D019887:Peanut Agglutinin / Q000032:analysis / Q000378:metabolism; D037121:Plant Lectins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D012680:Sensitivity and Specificity; D014909:Wheat Germ Agglutinins / Q000032:analysis / Q000378:metabolism
D000906:Antibodies; D006023:Glycoproteins; D019887:Peanut Agglutinin; D037121:Plant Lectins; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A