[18] Streptavidin-biotinylglycopeptide-lectin complex in detection of glycopeptides and determination of lectin specificity

Publisher Summary Streptavidin–biotinylglycopeptide–lectin complexes can be used in the detection of glycopeptides and the determination of lectin specificity at the picomole level. The method is based on the high affinity between streptavidin and biotin derivatives, 1, 2 and between lectins and their complementary sugars, 3 and on the ability of streptavidin to adhere to microtiter plate wells 4'5. For the detection of glycopeptides, each unknown (glyco) peptide [from a high-performance liquid chromatography (HPLC) run] is first biotinylated and complexed with streptavidin, each conjugate is then coated to the well of a titer plate, and lectin coupled to horseradish peroxidase (HRP) is added. After incubation, the wells are thoroughly washed and probed for the presence of the peroxidase. A positive test shows the presence of a glycan complementary to the lectin used.