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.
SEC-ICP-MS chromatogram of metal tagged antibody
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Acknowledgments
The cooperation between BAM and BOKU was financially supported by a publication fellowship of “Gesellschaft Deutscher Chemiker e.V.” The authors thank Christian Scheler (Proteomefactory, Berlin, Germany) for providing MeCAT reagents.
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Mueller, L., Mairinger, T., Hermann, G. et al. Characterization of metal-tagged antibodies used in ICP-MS-based immunoassays. Anal Bioanal Chem 406, 163–169 (2014). https://doi.org/10.1007/s00216-013-7416-x
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DOI: https://doi.org/10.1007/s00216-013-7416-x