Analytische Chemie
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Current trends in materials and life sciences are flanked by the need to push detection limits to single molecules or single cells, enable the characterization of increasingly complex matrices or sophisticated nanostructures, speed up the time of analysis, reduce instrument complexity and costs, and improve the reliability of data. This requires suitable analytical tools such as spectroscopic, separation and imaging techniques, mass spectrometry, and hyphenated techniques as well as sensors and their adaptation to application-specific challenges in the environmental, food, consumer product, health sector, nanotechnology, and bioanalysis. Increasing concerns about health threatening known or emerging pollutants in drinking water, consumer products, and food and about the safety of nanomaterials led to a new awareness of the importance of analytical sciences. Another important driver in this direction is the increasing demand by legislation, particularly in view of the 17 sustainable development goals by the United Nations addressing clean energy, industry, and innovation, sustainable cities, clean water, and responsible consumption and production. In this respect, also the development of analytical methods that enable the characterization of material flows in production processes and support recycling concepts of precious raw materials becomes more and more relevant. In the future, this will provide the basis for greener production in the chemical industry utilizing recycled or sustainable starting materials.
This makes analytical chemistry an essential player in terms of the circular economy helping to increase the sustainability of production processes. In the life sciences sector, products based on proteins, such as therapeutic and diagnostic antibodies, increase in importance. These increasingly biotechnologically produced functional biomolecules pose a high level of complexity of matrix and structural features that can be met only by highly advanced methods for separation, characterization, and detection. In addition, metrological traceability and target definition are still significant challenges for the future, particularly in the life sciences.
However, innovative reference materials as required for the health and food sector and the characterization of advanced materials can only be developed when suitable analytical protocols are available. The so-called reproducibility crisis in sciences underlines the importance of improved measures of quality control for all kinds of measurements and material characterization. This calls for thorough method validation concepts, suitable reference materials, and regular interlaboratory comparisons of measurements as well as better training of scientists in analytical sciences.
The important contribution of analytical sciences to these developments is highlighted by a broad collection of research papers, trend articles, and critical reviews from these different application fields. Special emphasis is dedicated to often-overlooked quality assurance and reference materials.
The mechanism of this system is based on kinetic competition. This biosensor consists of a monolithic glass column with a vast excess of immobilized hapten, which traps the fluorescently labeled antibody as long as no explosive is present. If the explosive 2,4,6-trinitrotoluene (TNT) is introduced some binding sites of the antibody will be blocked, which leads to an immediate breakthrough of the labeled protein. The fluorescence is detected by highly sensitive laser-induced fluorescence with a conventional CMOS camera. The system achieved limits of detection of approx.1 pM (1 ppt) of the fluorescent label and around 100 pM (20 ppt) of TNT. The total assay time is less than 8 minutes. A cross-reactivity test with 5000 pM solutions of pentaerythritol tetranitrate (PETN), 1,3,5-trinitroperhydro-1,3,5-triazine (RDX), and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) showed no cross reactivity.
Methods for the rapid and sensitive detection of target analytes are gaining importance in medical diagnostics and environmental monitoring, in the security, occupational health and safety as well as food sectors. Among all of the methods employed for rapid tests, lateral flow assays (LFAs) are the most commonly used, and hundreds of test kits based on this technique are available on the market. A major drawback is that most of these capture agents either indicate the analyte only indirectly, and in most cases a second binding agent able to bind directly or indirectly to the analytes is necessary (e.g., a secondary labeled antibody). Furthermore, in certain cases in which the (ultra)trace detection of an analyte is required, the traditional approach of a certain number of probe molecules being conjugated to a particular support is not sufficient. Therefore, novel concepts implementing steps of effective signal amplification are urgently required.
Keeping in mind these limitations, we thought that the sensitivity of these systems should be improvable through employment of gated reporter molecule-releasing hybrid nanoparticle materials on novel lateral flow devices. On one hand, the gated sensor material can produce a massive signal amplification, by releasing many reporter molecules only after chemical recognition of a few analyte molecules has taken place in an independent and separate step at the pore openings. On the other hand, the employment of tailored capture materials for the selective interaction with the released reporter molecules in a second arbitrary zone on the strip allows to concentrate or focus the latter for more efficient detection or to create selective multi-spot detection zones, which renders the simultaneous detection of several reporter molecules at the same time in multiplexed detection of various analytes possible.
For that purpose, we have prepared several stimuli-responsive materials for small-molecule sensing based on specific interactions between biomolecules such as antibodies with the corresponding analytes for the detection of certain explosives. In order to prepare these bio-capped materials, we have selected silica mesoporous nanoparticles (MSNs) as inorganic support due to their unique properties such as defined void structure, high inner surface area and flexible functionalization chemistry. These MSNs are loaded with a brightly fluorescent indicator dye, and the external surface is subsequently functionalized with suitable molecules able to interact with antibodies, efficiently inhibiting dye release. The opening protocol and delivery of the entrapped dye is reminiscent of a displacement reaction involving the presence of the target analyte, producing a displacement of the biomolecule and allowing the detection of the target analyte.
The presentation discusses general aspects of system design as well as analytical performance and highlights the integration into a lateral-flow assay, showing as an example the determination of the explosives TATP, TNT and PETN with fluorescence readout, in single-substance and multiplexing modes.
Phage display is used to find specific target binding peptides for polypropylene (PP) surfaces. PP is one of the most commonly used plastics in the world. Millions of tons are produced every year. PP binders are of particular interest because so far gluing or printing on PP is challenging due to its low surface energy. A phage display protocol for PP was developed followed by Next Generation DNA Sequencing of the whole phage library. Data analysis of millions of sequences yields promising peptide candidates which were synthesized as PEG conjugates. Fluorescence-based adsorption-elution-experiments show high adsorption on PP for several sequences.