title: Profile identification of disease-associated humoral antigens using AMIDA, a novel proteomics-based technology.
abstract: We describe AMIDA (autoantibody-mediated identification of antigens), a novel target identification technology based on the immunoprecipitation of disease-specific antigens by autologous serum antibodies followed by two-dimensional electrophoretic separation, and their identification via mass spectrometry. Twenty-seven potential carcinoma antigens were identified including proteins of hitherto unknown function. Validation of one of the identified antigens, cytokeratin 8, revealed its de novo expression in hyperplastic tissue, gradual overexpression with increasing malignancy, and ectopic localization on the cell surface. Furthermore, a strong prevalence of CK8-specific antibodies occurred in the serum of cancer patients already at early disease stages. In situ hybridization for one marker of unknown function, KIAA1273/TOB3, demonstrated its strong overexpression in head and neck carcinomas, thus making it a likely tumor antigen candidate. Eventually, AMIDA could foster significant improvements for the diagnosis and therapy of human diseases eliciting a humoral immune response, and allows for the rapid identification of new target molecules.
authors.affiliation: Department of Head and Neck Surgery, Ludwig-Maximilians University of Munich, Marchioninistr. 15, 81377 Munich, Germany. ogires@hno.med.uni-muenchen.de
authors.forename: O
authors.initials: O
authors.lastname: Gires
authors.forename: M
authors.initials: M
authors.lastname: Münz
authors.forename: M
authors.initials: M
authors.lastname: Schaffrik
authors.forename: C
authors.initials: C
authors.lastname: Kieu
authors.forename: J
authors.initials: J
authors.lastname: Rauch
authors.forename: M
authors.initials: M
authors.lastname: Ahlemann
authors.forename: D
authors.initials: D
authors.lastname: Eberle
authors.forename: B
authors.initials: B
authors.lastname: Mack
authors.forename: B
authors.initials: B
authors.lastname: Wollenberg
authors.forename: S
authors.initials: S
authors.lastname: Lang
authors.forename: T
authors.initials: T
authors.lastname: Hofmann
authors.forename: W
authors.initials: W
authors.lastname: Hammerschmidt
authors.forename: R
authors.initials: R
authors.lastname: Zeidler
mesh_terms: D000486:Allergy and Immunology; D000906:Antibodies / Q000737:chemistry; D000917:Antibody Formation; D000951:Antigens, Neoplasm / Q000737:chemistry; D002294:Carcinoma, Squamous Cell / Q000276:immunology; D045744:Cell Line, Tumor; D002469:Cell Separation; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007633:Keratins / Q000737:chemistry; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D011233:Precipitin Tests; D040901:Proteomics / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D000906:Antibodies; D000951:Antigens, Neoplasm; D007633:Keratins