Binding parameters were determined for four mouse monoclonal antibodies reacting with three antigens on the surface of fresh human ovarian carcinoma ascites cells, under nearly physiological conditions. The object of these experiments was to aid in the selection of the optimal monoclonal antibodies for intraperitoneal immunotherapy. The number of antigenic sites per cell, the effective equilibrium association constant (affinity) and the half-life for dissociation were: for Ab MH99, 1.2 x 10(6) sites/cell, (1.9-4.1) x 10(8) M-1, and 4 h; for Ab MX35, (3.2-4.1) x 10(5) sites/cell, (3.4-4.8) x 10(8) M-1, and greater than 10 h; and for Ab MW207, 1.3 x 10(5) sites/cell, (3.6-4.1) x 10(9) M-1, and 3.1 h, respectively. One of the antigens, MH99, is recognized by five different monoclonal antibodies, and competitive inhibition experiments demonstrated that two distinct determinants are present; this antigen is also recognized by the previously described Ab 17-1A. These binding data will aid the rational design of immunotherapy strategies.