CELL-CELL ADHERENCE AS A SELECTION METHOD FOR THE GENERATION OF ANTIMELANOMA MONOCLONAL-ANTIBODIES

The aim of the present study was to obtain monoclonal antibodies (mAbs ) recognising human melanoma-associated antigens after immunisation of BALB/c mice with a 70-150 kDa membrane fraction from melanoma tumour tissues. Screening of specific antibody- producing hybridomas was perf ormed using a novel cell-cell adherence method with the melanoma cell line M-14. Three mAbs of IgG1 isotype were selected: Mel-1, Mel-2 and Mel-3 which recognised the immunogen by ELISA and stained several mela noma cell lines positive in immunofluorescence. The molecular weight o f the antigen was studied by different methods; a 170-kDa band was ide ntified following immunoblotting of tumour lysate and a 72-kDa band wa s observed following immunoaffinity purification, Cell-cell adherence appears to be a reliable procedure for the generation of mAbs against native cellular antigens.