title: Multiplexed immunoassay for the rapid detection of anti-tumor-associated antigens antibodies.
abstract: TAAs (tumor-associated antigens) microarrays were designed to detect auto-antibodies directly in patient sera. Twelve different probes were chosen according to their described occurrence in cancer pathologies (Cyclin B1, Cyclin D1, Complement factor H, c-myc, IMP1, p53, p62, survivin, Her2/neu, Koc, NY-ESO-1 and PSA). Microarrays of these 12 proteins were immobilized within the nitrocellulose/cellulose acetate membrane of a 96-well filtering microtiter plate bottom. The captured auto-antibodies were detected using a staining approach based on alkaline phosphatase labeling. Thus, the presence of specific auto-antibodies in samples was visualized through the positive staining of the corresponding TAA spots. The TAA HiFi microarrays were shown to be able to capture specific purified anti-TAA antibodies. In real samples, 9 proteins from the 12 TAAs panel were shown to generate specific signal and 5 antigens (p53, NY-ESO-1, IMP1, cyclin B1 and c-myc) were shown to have interaction with more than 10% of the positive sera from cancer patients. This protein subpanel was proven to be able to detect 72.2% of the cancer patients tested (within a 34 panel of 18 patients and 16 healthy donors).
authors.affiliation: Equipe Génie Enzymatique, Membranes Biomimétiques et Assemblages Supramoléculaires, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université Lyon 1-CNRS 5246 ICBMS, Villeurbanne Cedex, France.
authors.forename: C
authors.initials: C
authors.lastname: Desmet
authors.forename: G C
authors.initials: GC
authors.lastname: Le Goff
authors.forename: J-C
authors.initials: JC
authors.lastname: Brès
authors.forename: D
authors.initials: D
authors.lastname: Rigal
authors.forename: L J
authors.initials: LJ
authors.lastname: Blum
authors.forename: C A
authors.initials: CA
authors.lastname: Marquette
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000097:blood / Q000276:immunology*; D001323:Autoantibodies / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods
chemical_list: D000951:Antigens, Neoplasm; D001323:Autoantibodies; D000469:Alkaline Phosphatase