Biotech drugs are poorly absorbed, highly susceptible to loss of activity, and require formulations with protective effects. Their incorporation into nanospheres is a promising approach since colloidal formulations were shown to be absorbed. Due to rather low absorption rates the surface engineering with absorption enhancing agents is a current challenge. Thus, an optimized protocol relying on carbodiimide-mediated covalent binding of biorecognitive ligands to the surface of biodegradable and biocompatible poly(D,L-lactic-co-glycolic)acid (PLGA) nanospheres was established. Important parameters such as retainment of particle size distribution and biorecognitive characteristics of the ligand as well as storage stability were considered. As exemplified by nanoparticles grafted with wheat germ agglutinin (WGA) as a carbohydrate binding ligand and Caco-2 single cells, flow cytometry as well as confocal laser scanning microscopy revealed improved cytoassociation of the nanoscaled carriers as compared to the unmodified ones. As detected by flow cytometry, the binding of WGA-decorated nanospheres considerably increases the roughness of the cell surface with increasing ligand density. Additionally, the biorecognitive nanoparticles were not toxic in the Caco-2 model. The protocol presented for surface modification of nanospheres promises high versatility of application in search for biorecognitive ligands enhancing the cytoadhesion, cytoinvasion as well as transcellular transport of colloidal carriers.