GENERATION AND SELECTION OF MONOCLONAL-ANTIBODIES, SINGLE-CHAIN FV AND ANTIBODY FUSION PHAGE SPECIFIC FOR HUMAN MELANOMA-ASSOCIATED ANTIGENS

A panel of 13 murine monoclonal antibodies (mAbs) recognizing antigens on human melanoma cells but not on melanocytes was generated, Two mAb s (LHM3 and LHM5) stained sections of melanoma but not normal tissues, mAbs LHM2 and LHM8 stained only a minority of normal tissues, The mAb s differed further in their staining patterns on melanoma cell lines H MB2, DX3 and SK23 in FAGS, The mAbs recognize antigens of 34, 38, 57, 94, 190-200 and >200 kD. One mAb each bound to each of the antigens HL A DR (LHM4) and high molecular weight proteoglycan (LHM2). The high mo lecular weight proteoglycan-specific mAb was used to construct a singl e-chain Fv (scFv) antibody fragment and an antibody fusion phage in Es cherichia coil, Both the scFv and the fusion phage were shown to bind specifically to melanoma cells, A method for the selection of melanoma cell-binding phages from phage libraries is described.