Generally, the rate of mAb and ADC internalization is influenced by a number of parameters, including epitope selectivity, binding affinity, and intracellular transportation. A cell surface receptor offers many epitopes for mAbs binding and different antibody-epitope interactions reveal a wide distribution of internalization kinetics. Thus, the selection of the antibody-epitope pair with the most suitable internalization kinetics is an important factor in ADC development. Additionally, earlier studies have revealed an intricate correlation between antibody internalization, tissue penetration, and binding affinity:

1.antibodies with moderate affinity exhibited the highest levels of tumor accumulation, indicating good tumor penetration, whereas high-affinity antibodies usually exhibited a low level of tumor accumulation.

2.the internalization kinetics is related to binding affinity, with high-affinity antibodies exhibited greater internalization potential comparing to low-affinity antibodies.