TY - JOUR A1 - Müller, Larissa A1 - Mairinger, T. A1 - Hermann, G. A1 - Koellensperger, G. A1 - Hann, S. T1 - Characterization of metal-tagged antibodies used in ICP-MS-based immunoassays JF - Analytical and bioanalytical chemistry N2 - 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. KW - Bioanalytical methods KW - Mass spectrometry KW - ICP-MS KW - Antibody tagging KW - Antibodies KW - Immunoassay KW - Maleimide DOTA KW - Tags PY - 2014 DO - https://doi.org/10.1007/s00216-013-7416-x SN - 1618-2642 SN - 1618-2650 VL - 406 IS - 1 SP - 163 EP - 169 PB - Springer CY - Berlin AN - OPUS4-30037 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Völzke, Jule L. A1 - Hodjat Shamami, Parya A1 - Gawlitza, Kornelia A1 - Feldmann, Ines A1 - Zimathies, Annett A1 - Meyer, Klas A1 - Weller, Michael G. T1 - High-Purity Corundum as Support for Affinity Extractions from Complex Samples JF - Separations N2 - Nonporous corundum powder, known as an abrasive material in the industry, was functionalized covalently with protein binders to isolate and enrich specific proteins from complex matrices. The materials based on corundum were characterized by TEM, ESEM, BET, DLS, EDS, and zeta potential measurements. The strong Al-O-P bonds between the corundum surface and amino phosphonic acids were used to introduce functional groups for further conjugations. The common crosslinker glutaraldehyde was compared with a hyperbranched polyglycerol (PG) of around 10 kDa. The latter was oxidized with periodate to generate aldehyde groups that can covalently react with the amines of the surface and the amino groups from the protein via a reductive amination process. The amount of bound protein was quantified via aromatic amino acid analysis (AAAA). This work shows that oxidized polyglycerol can be used as an alternative to glutaraldehyde. With polyglycerol, more of the model protein bovine serum albumin (BSA) could be attached to the surface under the same conditions, and lower non-specific binding (NSB) was observed. As a proof of concept, IgG was extracted with protein A from crude human plasma. The purity of the product was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A binding capacity of 1.8 mg IgG per gram of corundum powder was achieved. The advantages of corundum include the very low price, extremely high physical and chemical stability, pressure resistance, favorable binding kinetics, convenient handling, and flexible application. KW - Corundum KW - Sapphire KW - Affinity chromatography KW - Antibodies KW - Self-assembled monolayers (SAM) KW - Polyglycerol KW - Dendrimer KW - Nonspecific binding (NSB) KW - Purification KW - Solid-phase extraction (SPE) PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-559342 DO - https://doi.org/10.3390/separations9090252 VL - 9 IS - 9 SP - 1 EP - 13 PB - MDPI CY - Basel AN - OPUS4-55934 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -