Uptake Kinetics of Monoclonal Antibodies by Human Malignant Melanoma Multicell Spheroids¹

Detailed uptake kinetics by multicell spheroids of three tumor associated monoclonal antibodies was investigated. The spheroids were established from a human melanoma cell line and the human colon adenocarcinoma cell line HT29 as in vitro models of poorly vascularized micrometastases in vivo. The selected antibodies 96.5, 140.240, and OST15 showed a wide range of reactivity against the melanoma cell but they all had negligible binding with the colon cancer cell. Uptake of the antibodies by small spheroids (about 300 µm diameter) was generally sigmoidal in shape with respect to incubation time, and amount of uptake followed the same trend of immunoreactivity of the antibodies with single cells. The correlation was weaker for spheroids with diameter greater than 500 µm presumably due to the increasing size of the necrotic core. By varying the concentration of the antibodies in the incubation medium from tracer dose (0.2 µg/ml) to a higher dose (3 µg/ml), negligible changes in the amount of antibodies bound with their target spheroids were observed. Nonspecific binding between antibodies and spheroids, however, resulted in proportional increase in uptake.