Reactivity Spectrum of 30 Monoclonal Antimelanoma Antibodies to a Panel of 28 Melanoma and Control Cell Lines

Abstract

Thirty monoclonal antibodies from eight laboratories exchanged after the First Workshop on Monoclonal Antibodies to Human Melanoma held in March 1981 at NIH were tested in an antibody-binding radioimmunoassay using a panel of 28 different cell lines. This panel included 12 melanomas, three neuroblastomas, four gliomas, one retinoblastoma, four colon carcinomas, one lung carcinoma, one cervical carcinoma, one endometrial carcinoma, and one breast carcinoma. The reactivity pattern of the 30 monoclonal antibodies tested showed that none of them were directed against antigens strictly restricted to melanoma, but that several of them recognize antigenic structures preferentially expressed on melanoma cells. A large number of antibodies were found to crossreact with gliomas and neuroblastomas. Thus, they seem to recognize neuroectoderm associated differentiation antigens. Four monoclonal antibodies produced in our laboratory were further studied for the immunohistological localization of melanoma associated antigens on fresh tumor material. In a three-layer biotin-avidin-peroxidase system each antibody showed a different staining pattern with the tumor cells, suggesting that they were directed against different antigens.

Monoclonal antibodies that prevent adhesion of B 16 melanoma cells and reduce metastases in mice: crossreaction with human tumor cells.

“…Our data from this and the preceding work (12) A series of human melanoma-associated antigens have recently been identified in other laboratories by means of both poly-and monoclonal antibodies (13,(21)(22)(23)(24)(25)(26)(27)(28)(29)(30) chem-ically (e.g., CSG 120/7 carcinoma), and virally (e.g., SV40-3T3 and RSV-CEF) transformed cells and they are not species specific. Our antigens are clearly distinct from p53, however, because they all seem to be surface adhesion molecules.…”

Heterogenous expression of cell-surface antigens in normal epithelia and their tumours, revealed by monoclonal antibodies

“…Colon (Arends et al, 1983;Daar & Fabre, 1982;Finan et al, 1982) and lung (Wagenaar et al, 1984) tumours also show heterogeneity. In melanoma antibodies shown to stain heterogeneously include Me4-TB7, C13-C6 and Nu4B (Carrel et al, 1982;Thompson et al, 1982) as well as antibodies to HLA-DR as discussed below. The antibody SSEA-1 shows heterogenous staining of colon, stomach and kidney carcinoma, although breast carcinoma was almost homogeneously positive (Fox et al, 1983).…”

Development and validation of a cell-based fluorescent method for measuring antibody affinity

“…Development, production, and purification of the Mel-14 monoclonal antibody targeting human chondroitin sulfate proteoglycan 4 (CSGP4), the D2C7 monoclonal antibody targeting human wild-type epidermal growth factor receptor (EGFRwt) and human mutant EGFR variant III (EGFRvIII), and the NZ-1 monoclonal antibody targeting human podoplanin have been previously described [5-7]. The purity of Mel-14 (mouse IgG2a), D2C7 (mouse IgG1), and NZ-1 (Rat IgG2a) were determined to be greater than 90% by SDS-PAGE (data not shown).…”