Fig. 1.

Experimental strategy. (A) The approach followed for the selection of specific anti-Tn mAbs is schematically illustrated. Tn conjugated with cBSA was used for mice immunization, whereas Tn conjugated with biotinylated alginate was used for mouse serum titration and for the screening of anti-Tn clones. The selected hybridomas were further tested by ELISA and immunofluorescence on a highly expressing Tn breast cancer cell line (MCF7) and finally, after Qdot conjugation, were used for in vivo imaging. (B) Schematic representation of Tn-conjugates. The three compounds were obtained starting from pure Tn antigen, synthesized in seven steps from 2-azido-2-deoxy-1,3,4,6-tetra-O-acetyl galactose, exploiting the trichloroacetimidate strategy. All the derivatives were obtained applying the same carbodiimide chemistry: EDC and NHS were used as activating agents for the carboxylic group in the MES buffer, pH 6.0. The products were fully characterized before further utilization.