title: Reliability of the nanopheres-DNA immunization technology to produce polyclonal antibodies directed against human neogenic proteins.
abstract: The molecular domestication of several DNA transposons that occurred during the evolution of the mammalian lineage, has led to the emergence of at least 43 genes, known as neogenes. To date, the limited availability of efficient commercial antibodies directed against most of their protein isoforms hampers investigation of their expression in vitro and in situ. Since immunization protocols using peptides or recombinant proteins have revealed that it is difficult to recover antibodies, we planned to produce antisera in mice using a new technique of nanopheres/DNA immunization, the ICANtibodies™ technology. Here, we investigate the possibilities of obtaining polyclonal antibodies for 24 proteins or protein domains using this immunization strategy. We successfully obtained 13 antisera that were able to detect neogenic proteins by Western blotting and ELISA in protein extracts of transiently-transfected cells and various cancer cell lines, plus another two that only detected the in ELISA and in in situ hybridizations. The features required for the production of these antibodies are analyzed and discussed, and examples are given of the advantages they offer for the study of neogenic proteins.
authors.affiliation: UMR CNRS 7292, GICC, Université François Rabelais, 37200, Tours, France.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Arnaoty
authors.forename: Valérie
authors.initials: V
authors.lastname: Gouilleux-Gruart
authors.forename: Sophie
authors.initials: S
authors.lastname: Casteret
authors.forename: Bruno
authors.initials: B
authors.lastname: Pitard
authors.forename: Yves
authors.initials: Y
authors.lastname: Bigot
authors.forename: Thierry
authors.initials: T
authors.lastname: Lecomte
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D000917:Antibody Formation; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D004251:DNA Transposable Elements / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007114:Immunization / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D054118:Nanospheres / Q000737:chemistry; D011506:Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D000906:Antibodies; D000941:Antigens; D004251:DNA Transposable Elements; D007106:Immune Sera; D011506:Proteins