Monoclonal anti-melanoma antibodies IKH-1 and IKH-2 which work on formalin-fixed, paraffin embedded tissues: characterization, clinical trials and comparative studies with HMB-45.
Mouse monoclonal antibodies (MoAbs), IKH-1 and IKH-2, were produced against cloned human melanoma cells, KHm-6, which were cultured with 12-O-tetradecanoylphorbol-13-acetate (TPA) and processed by formalin fixation and alcohol dehydration (FFAD). According to the biochemical analysis, antigenic substances which reacted with IKH-1 were 34.0-60.0 kDa glycoproteins, and those which reacted with IKH-2 were 33.5, 34.5 and 36.0 kDa glycoproteins. Immunoelectron microscopy revealed that reaction products of IKH-1 were seen in some membranous vesicles, premelanosomes and cell membrane of TPA-treated KHm-6 cells, while IKH-2 recognized only premelanosomal structures. Immunohistochemical tests revealed that IKH-1 and IKH-2 have a high sensitivity (94.0% and 85.0%, respectively) to formalin-fixed, paraffin-embedded (FFPE) tissue sections of malignant melanomas. IKH-1 had a high specificity and IKH-2 and 100% specificity to FFPE tissue sections of melanocytic lesions.
Ishihara
M
M
Department of Dermatology, Wayne State University School of Medicine, Detroit, MI.
Hashimoto
K
K
DiGregorio
F M
FM
Setoyama
M
M
Richards
J
J
Pietruk
T
T
Zarbo
R J
RJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000276:immunology; D000951:Antigens, Neoplasm / Q000032:analysis / Q000276:immunology; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000276:immunology; D002471:Cell Transformation, Neoplastic / Q000187:drug effects / Q000473:pathology; D002986:Clinical Trials as Topic; D005557:Formaldehyde; D006801:Humans; D006825:Hybridomas / Q000737:chemistry / Q000276:immunology / Q000473:pathology; D007150:Immunohistochemistry; D008545:Melanoma / Q000737:chemistry / Q000276:immunology* / Q000473:pathology; D008546:Melanoma, Experimental / Q000737:chemistry / Q000276:immunology / Q000473:pathology; D051379:Mice; D016253:Microscopy, Immunoelectron; D010232:Paraffin; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D016707:Tissue Fixation / Q000379:methods; D014407:Tumor Cells, Cultured / Q000276:immunology / Q000473:pathology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D005557:Formaldehyde; D010232:Paraffin; D013755:Tetradecanoylphorbol Acetate