title: Characterization of metal-tagged antibodies used in ICP-MS-based immunoassays.
abstract: A detailed characterization of metal-tagged antibodies is the prerequisite for the implementation of quantitative concepts in inductively coupled plasma-mass spectrometry (ICP-MS)-based bioanalysis or future medical diagnosis. In this paper, the common modification with bifunctional ligands containing maleimide residues as a reactive group was investigated in detail via size exclusion chromatography (SEC)-ICP-MS and liquid chromatography-time-of-flight (LC-TOF)-MS to determine the preservation of the antibody structure after tagging. Mouse monoclonal IgG modified with metal-coded tags (MeCATs) was used as a model system. Several antibody fragments were identified carrying different numbers of metal tags. In a second step, a functionality test was performed with isolated fragments where the antigen specificity was tested in a dot blot immunoassay.
authors.affiliation: BAM Federal Institute for Materials Research and Testing, Richard-Willstaetter-Straße 11, 12489, Berlin, Germany, larissa.mueller@bam.de.
authors.forename: L
authors.initials: L
authors.lastname: Mueller
authors.forename: T
authors.initials: T
authors.lastname: Mairinger
authors.forename: G
authors.initials: G
authors.lastname: Hermann
authors.forename: G
authors.initials: G
authors.lastname: Koellensperger
authors.forename: S
authors.initials: S
authors.lastname: Hann
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007074:Immunoglobulin G / Q000737:chemistry*; D008301:Maleimides / Q000737:chemistry*; D051379:Mice; D009211:Myoglobin / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013194:Staining and Labeling / Q000379:methods; D013725:Terbium / Q000737:chemistry*
chemical_list: D000941:Antigens; D007074:Immunoglobulin G; D008301:Maleimides; D009211:Myoglobin; D013725:Terbium