Binding of biotinylated legume seed lectins with glycoproteins in blotted receptor-analogs: influence of incubation pH

SDS–PAGE and Western blotting of lectin inhibitors (bovine and porcine plasma powder, whole egg powder and fetuin) was performed and blots were incubated with several biotinylated lectins commercially available (Phaseolus vulgaris E, Pisum sativum, Lens culinaris, Vicia faba). In order to study the effect of pH on the binding of the lectins to glycoproteins present in the inhibitors, identical blots were incubated in buffers with different pH values, i.e. 3–7, respectively. Binding capacity of lectins to the glycoproteins in the inhibitors was very dependent on pH condition during incubation: for all the lectins involved in this experiment, pH values lower than 4 inhibited the binding process considerably. Results are discussed taking into account the low pH values prevailing in the stomach of pigs.