GENERATION OF HUMAN MONOCLONAL-ANTIBODIES AGAINST GANGLIOSIDE ANTIGENS AND THEIR APPLICATIONS IN THE DIAGNOSIS AND THERAPY OF CANCER

Different approaches to generating human monoclonal antibodies (MAbs) against tumor-associated ganglioside antigens have been carried out in several laboratories, A specific goal addressed by our laboratory is to produce human MAbs to several ganglioside antigens of relevance as therapeutic targets, such as the GM2, GD2, GD3 and GM3 gangliosides in melanoma. In vitro immunization of human B lymphocytes from normal do nors was performed using liposomes containing gangliosides as the immu nizing antigen combined with either complete tetanus toroid or a synth etic peptide corresponding to a T helper epitope to stimulate in vitro immunization, Specific human anti-ganglioside antibodies were obtaine d, indicating that the antibody response found in vitro was antigen-dr iven, To overcome the widely reported problems concerning stability of immunoglobulin production by the antibody-secreting cell lines, a met hod of positive selection using GM3-coated magnetic beads has been dev eloped in order to rescue unstable clones, Development of new methods to reproducibly generate ganglioside-specific human MAbs will amplify the possibilities for diagnostic and therapeutic applications.