Cell-cell adherence as a selection method for the generation of anti-melanoma 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 performed 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 melanoma cell lines positive in immunofluorescence. The molecular weight of the antigen was studied by different methods; a 170-kDa band was identified following immunoblotting of tumour lysate and a 72-kDa band was observed following immunoaffinity purification. Cell-cell adherence appears to be a reliable procedure for the generation of mAbs against native cellular antigens.
Barral
A M
AM
Clinical Immunology Group, National Institute of Oncology and Radiobiology, Havana, Cuba. AnnBa@MCB.LIU.SE
Fernández
A
A
Faxas
M E
ME
Pérez
X
X
García
C A
CA
Rosén
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000912:Antibodies, Neoplasm / Q000096:biosynthesis*; D000951:Antigens, Neoplasm / Q000276:immunology / Q000302:isolation & purification; D002448:Cell Adhesion / Q000276:immunology; D002846:Chromatography, Affinity; D006801:Humans; D006825:Hybridomas / Q000737:chemistry; D007150:Immunohistochemistry; D008545:Melanoma / Q000276:immunology*; D058950:Melanoma-Specific Antigens; D051379:Mice; D008807:Mice, Inbred BALB C; D009363:Neoplasm Proteins / Q000276:immunology; D010954:Plasmacytoma; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins