TY - JOUR A1 - Carvalho, Ana Margarida A1 - Véliz Montes, Cinthya A1 - Schneider, Rudolf A1 - Madder, A. T1 - An anti-caffeine antibody−oligonucleotide conjugate for DNA-directed immobilization in environmental immunoarrays N2 - The development of fast and cheap highthroughput platforms for the detection of environmental contaminants is of particular importance to understand the human-related impact on the environment. The application of DNA-directed immobilization (DDI) of IgG molecules is currently limited to the clinical diagnostics scenario, possibly because of the high costs of production of such addressable platforms. We here describe the efficient and specific hybridization of an antibody−oligonucleotide conjugate to a short 12-mer capture probe. The specific antibody used is a monoclonal antibody against caffeine, a stimulant and important anthropogenic marker. With this work, we hope to contribute to broadening the application potential of DDI to environmental markers in order to develop cheaper and more stable high-throughput screening platforms for standard Routine analysis of pollutants in a variety of complex matrices. KW - Oligo KW - Antikörperkonjugate KW - Antikörper KW - Biokonjugation KW - Koffein KW - Array PY - 2018 DO - https://doi.org/10.1021/acs.langmuir.8b01347 SN - 0743-7463 VL - 34 IS - 49 SP - 14834 EP - 14841 PB - American Chemical Society CY - Hoboken, New Jersey, USA AN - OPUS4-46982 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Carvalho, Margarida T1 - Addressable Immunoarrays: Protein-Nucleic Acid Conjugate Production N2 - Immunoassays are analytical methods used to track both clinical and environmental parameters. Antibodies or other proteins with similar recognizing activity, are employed, often immobilized onto a modified surface. Addressable microarrays are based on single-stranded DNA oligonucleotides which are normally used to detect aptamers or relevant gene sequences. The use of antibody-oligonucleotide conjugates allows non-directed antibody immobilization from an immunoassay to be converted into DNA hybridization events on the array. Consequently, the diagnostics platform is multiplexed and addressable. Figure 1 is a model representation of the whole biosensor construct. In brief, a surface is coated with streptavidin, and decorated with biotin-modified oligonucleotides of a controlled size and known sequence. These oligonucleotides on the surface also contain a furan motif, which upon irradiation and in the presence of a photosensitizer, is oxidized via single oxygen to a reactive intermediate which crosslinks the immediate opposing base when hybridized with its complementary strand [1]. Antibody-oligonucleotides conjugates still display several concerns due to their heterogeneicity, difficulty of characterization and high price [2]. The being the case, we aim at developing stable, robust, reproducible and well characterized quimeras for the application on te above described immunoarray. Mild chemical conditions are crucial for the antibody stability, therefore bivalent crosslinkers have been employed [2]. Even though the use of these bilinkers is standard for other substrate conjugation (i.e., enzymes, drug payloads, etc.), there is no literature available on the conjugation of small oligonucleotides (< 30 mer) to antibodies using this method. Several protein-nucleic acid conjugates have been therefore developed, and successfully characterized using MALDI-ToF and gel electrophoresis techniques. T2 - SO2S - The singlet Oxygen Strategy - Final Network Meeting CY - Gent, Belgium DA - 14.06.2016 KW - Antibodies KW - Immunoarrays KW - Nucleic acids PY - 2016 AN - OPUS4-36680 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -