Development and characterization of a murine monoclonal antibody against phaeomelanin and its precursor 5-S-cysteinyldopa.
This study reports our success in developing a murine monoclonal antibody (mAb) against phaeomelanin and its major precursor, 5-S-cysteinyldopa (5-S-CD). A competitive enzyme-linked immunosorbent assay was developed and demonstrated that the new mAb, designated as 6D4 (IgG1, kappa), reacted with both 5-S-CD and phaeomelanin, but not with eumelanin. The concentrations required for 50% inhibition of 5-S-CD and phaeomelanin were 65 ng/ml and 95 ng/ml respectively. The minimal amount of 5-S-CD and phaeomelanin which could be detected by mAb 6D4 was approximately 5 and 6 ng/ml, respectively. The immunohistochemical assay indicated that the antigenic epitope(s) recognized by mAb 6D4 was localized in the cytoplasm of certain types of melanocytic tumours, such as superficial spreading melanoma.
Liu
J
J
Division of Dermatology and Cutaneous Sciences, University of Alberta, Edmonton, Canada.
Jimbow
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001667:Binding, Competitive; D003548:Cysteinyldopa / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification; D008543:Melanins / Q000276:immunology*; D008545:Melanoma / Q000276:immunology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D009369:Neoplasms / Q000276:immunology / Q000473:pathology; D009508:Nevus, Pigmented / Q000276:immunology / Q000473:pathology; D051381:Rats; D012878:Skin Neoplasms / Q000276:immunology / Q000473:pathology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008543:Melanins; C016042:phaeomelanin; C041877:eumelanin; D003548:Cysteinyldopa