Targeting of carbonic anhydrase IX-positive cancer cells by glycine-coated superparamagnetic nanoparticles

Antibody-modified magnetic nanoparticles were prepared to study their cellular uptake in 3D multicellular spheroidal cell cultures. For this purpose, carbonic anhydrase IX specific monoclonal antibody VII/20 was selected to conjugate on the surface of positively charged glycine coated magnetic nanoparticles in a form of a stable magnetic fluid. In this work, glycine-functionalized magnetic nanoparticles were characterized by different methods. X-ray photoelectron analysis confirmed the binding of glycine to the magnetic nanoparticles, and quantification of the glycine coating on the surface of the magnetic nanoparticles was conducted by thermogravimetric analysis. The optimal weight ratio of glycine to magnetic nanoparticles was determined to be 5 showing good colloid stability due to the high surface charge density of protonated glycine coating shown by the great zeta potential (⁓40 mV). The antibody conjugation to the functionalized magnetic nanoparticles was performed at an antibody to magnetic nanoparticles weight ratio equal to 0.5. Applications of antibody-modified magnetic nanoparticles in cancer therapy rely on their ability to specifically target cancer tissues and enter the tumour intracellular space. Here, we show that antibody coupled nanoparticle internalization was triggered by selective binding to tumour cells expressing hypoxic marker carbonic anhydrase IX. Moreover, our results confirmed specific penetration of conjugated nanoparticles into the tumour cell spheroids.