Cellular immuno-PCR. Detection of a carbohydrate tumor marker.
Carbohydrate tumor-antigens are important tumor markers for diagnosis and functional characteristics of human cancer cells. Detection of these carbohydrate tumor antigens on metastatic cancer cells in blood is a difficult task. We developed a highly sensitive method to detect a cell surface carbohydrate antigen using a hybrid technology referred to as cellular immuno-PCR. This technique uses the human monoclonal antibody (HumAb) L612, specific to a tumor-related antigen (ganglioside) GM3 that is expressed on the cell surface of human tumor cells and not normal cells. L612 coupled to a DNA oligonucleotide for exponential amplification by DNA polymerase chain reaction (PCR) can be used to enhance the detection signal. The DNA-HumAb conjugate was assessed for detection of a small number of human cancer cells after PCR amplification and Southern blot analysis. To assess the assay specificity human melanoma and other cancer cell lines, as well as healthy donor and melanoma patients, bloods were assessed. Cellular immuno-PCR requires < 1 ng/ml DNA-HumAb complex and was shown to have a detection level of < 10 cells in titration studies in which melanoma cells were diluted in 2 million healthy donor peripheral blood lymphocytes. The assay was shown to be very sensitive and could detect low levels of GM3 antigen expression by tumor cells. This novel approach for detecting a carbohydrate tumor antigen on tumor cells in blood provides a potential useful clinicopathological assay.
Zhang
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Department of Biotechnology Sciences, John Wayne Cancer Institute, Santa Monica, California 90404, USA.
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D014408:Biomarkers, Tumor / Q000097:blood / Q000276:immunology*; D019921:Biotinylation; D015139:Blotting, Southern; D002241:Carbohydrates / Q000276:immunology*; D006801:Humans; D007118:Immunoassay; D007129:Immunoglobulin Gm Allotypes / Q000097:blood / Q000187:drug effects / Q000276:immunology*; D008545:Melanoma / Q000097:blood / Q000276:immunology*; D009439:Neuraminidase / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D014408:Biomarkers, Tumor; D002241:Carbohydrates; D007129:Immunoglobulin Gm Allotypes; D009439:Neuraminidase