Tailor-made recombinant prokaryotic lectins for characterisation of glycoproteins

Development of biosimilars is costly, where glycan analysis a significant constraint on time and money. This paper provides an in-depth characterisation several novel recombinant prokaryotic lectins (RPLs), developed through directed evolution, displaying specific binding activities to ?-mannose, ?-galactose, fucose sialic acid residues, tested against major biosimilar targets. The all was performed employing the principles bio-layer interferometry (BLI), with help streptavidin-coated sensor biotinylated lectins. activity RPLs specificity broad range glycoproteins glycoconjugates were evaluated compared those equivalent plant-derived While exhibiting better or similar specificity, displayed significantly in cases. mechanisms are explained particular focus role hydrogen bonding plays change for galactose lectin. Furthermore, different sets their plant equivalents assayed glycoprotein targets evaluate analytical parameters lectin-glycoprotein interaction. obtained LoDs reached by lower than counterparts apart from one, RPL:PL LoD ratios 0.8, 2.5, 14.2 380 fucose, ?-galactose acid, respectively. Such enhancement shows applicability protein purification as bioanalytical tools biosensor development.