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Item type:Publication, Access status: without files , Automated Laser-Transfer Synthesis of High-Density Microarrays for Infectious Disease Screening(2022-06-01); ; ; Laser-induced forward transfer (LIFT) is a rapid laser-patterning technique for high-throughput combinatorial synthesis directly on glass slides. A lack of automation and precision limits LIFT applications to simple proof-of-concept syntheses of fewer than 100 compounds. Here, an automated synthesis instrument is reported that combines laser transfer and robotics for parallel synthesis in a microarray format with up to 10 000 individual reactions cm<sup>−</sup><sup>2</sup>. An optimized pipeline for amide bond formation is the basis for preparing complex peptide microarrays with thousands of different sequences in high yield with high reproducibility. The resulting peptide arrays are of higher quality than commercial peptide arrays. More than 4800 15-residue peptides resembling the entire Ebola virus proteome on a microarray are synthesized to study the antibody response of an Ebola virus infection survivor. Known and unknown epitopes that serve now as a basis for Ebola diagnostic development are identified. The versatility and precision of the synthesizer is demonstrated by in situ synthesis of fluorescent molecules via Schiff base reaction and multi-step patterning of precisely definable amounts of fluorophores. This automated laser transfer synthesis approach opens new avenues for high-throughput chemical synthesis and biological screening.Publicationtype: Journal Article8 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , On-Chip Neo-Glycopeptide Synthesis for Multivalent Glycan Presentation(2020-08-06); ; ; ; Single glycan–protein interactions are often weak, such that glycan binding partners commonly utilize multiple, spatially defined binding sites to enhance binding avidity and specificity. Current array technologies usually neglect defined multivalent display. Laser-based array synthesis technology allows for flexible and rapid on-surface synthesis of different peptides. By combining this technique with click chemistry, neo-glycopeptides were produced directly on a functionalized glass slide in the microarray format. Density and spatial distribution of carbohydrates can be tuned, resulting in well-defined glycan structures for multivalent display. The two lectins concanavalin A and langerin were probed with different glycans on multivalent scaffolds, revealing strong spacing-, density-, and ligand-dependent binding. In addition, we could also measure the surface dissociation constant. This approach allows for a rapid generation, screening, and optimization of a multitude of multivalent scaffolds for glycan binding.Publicationtype: Journal Article6 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Laser-induced forward transfer of soft material nanolayers with millisecond pulses shows contact-based material deposition(2020-04-01); ; ; ; In this work, we present a qualitative and quantitative experimental analysis, as well as a numerical model, of a novel variant of the laser-induced forward transfer, which uses millisecond laser pulses. In this process, soft material nanolayer spots are transferred from a donor slide, which is coated with the soft material layer, to an acceptor slide via laser irradiation. This method offers a highly flexible material transfer to perform high-throughput combinatorial chemistry for the generation of biomolecule arrays. For the first time, we show visual evidence that the main transfer mechanism is contact-based, due to thermal surface expansion of the donor layer. Thus, the process is different from the many known variants of laser-induced forward transfer. We characterize the maximum axial surface expansion in relation to laser power and pulse duration. On this basis, we derive a numerical model that approximates the axial surface expansion within measurement tolerances. Finally, we analyze the topology of the transferred soft material nanolayer spots by fluorescence imaging and vertical scanning interferometry to determine width, height, and shape of the transferred material. Concluding from this experimental and numerical data, we can now predict the amount of transferred material in this process.Publicationtype: Journal Article9 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Combinatorial Synthesis of Peptoid Arrays via Laser-Based Stacking of Multiple Polymer Nanolayers(2019-03-01); 8 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Combinatorial Synthesis of Macromolecular Arrays by Microchannel Cantilever Spotting (µCS)(2018-08-02); ; Surface-bound microarrays of multiple oligo- and macromolecules (e.g., peptides, DNA) offer versatile options in biomedical applications like drug screening, DNA analysis, or medical diagnostics. Combinatorial syntheses of these molecules in situ can save significant resources in regard to processing time and material use. Furthermore, high feature densities are needed to enable high-throughput and low sample volumes as generally regarded in combinatorial chemistry. Here, a scanning-probe-lithography-based approach for the combinatorial in situ synthesis of macromolecules is presented in microarray format. Feature sizes below 40 µm allow for the creation of high-density arrays with feature densities of 62 500 features per cm<sup>2</sup>. To demonstrate feasibility of this approach for biomedical applications, a multiplexed array of functional protein tags (HA- and FLAG-tag) is synthesized, and selective binding of respective epitope recognizing antibodies is shown. This approach uses only small amounts of base chemicals for synthesis and can be further parallelized, therefore, opening up a route to flexible, highly dense, and cost-effective microarrays.Publicationtype: Journal Article3 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , A Trifunctional Linker for Purified 3D Assembled Peptide Structure Arrays(2018-02-01); ; ; 5 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Development of a poly(dimethylacrylamide) based matrix material for solid phase high density peptide array synthesis employing a laser based material transfer(2016-12-15); ; ; ; von Bojnicic-Kninski, Clemens M.Poly(dimethylacrylamide) (PDMA) based matrix materials were developed for laser-based in situ solid phase peptide synthesis to produce high density arrays. In this specific array synthesis approach, amino acid derivatives are embedded into a matrix material, serving as a “solid” solvent material at room temperature. Then, a laser pulse transfers this mixture to the target position on a synthesis slide, where the peptide array is synthesized. Upon heating above the glass transition temperature of the matrix material, it softens, allowing diffusion of the amino acid derivatives to the synthesis surface and serving as a solvent for peptide bond formation. Here, we synthesized PDMA six-arm star polymers, offering the desired matrix material properties, using atom transfer radical polymerization. With the synthesized polymers as matrix material, we structured and synthesized arrays with combinatorial laser transfer. With densities of up to 20,000 peptide spots per cm <sup>2</sup> , the resolution could be increased compared to the commercially available standard matrix material. Time-of-Flight Secondary Ion Mass Spectrometry experiments revealed the penetration behavior of an amino acid derivative into the prepared acceptor synthesis surface and the effectiveness of the washing protocols.Publicationtype: Journal Article2 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , Selective Functionalization of Microstructured Surfaces by Laser-Assisted Particle TransferMicrocavity arrays represent millions of different reaction compartments to screen, for example, molecular interactions, exogenous factors for cells or enzymatic activity. A novel method is presented to selectively synthesize different compounds in arrays of microcavities with up to 1 000 000 cavities per cm<sup>2</sup>. In this approach, polymer microparticles with embedded pre-activated monomers are selectively transferred into microcavities with laser radiation. After particle patterning, heating of the particle matrix simultaneously leads to diffusion and coupling of the monomers inside each microcavity separately. This method exhibits flexibility, not only in the choice of compounds, but also in the choice of particle matrix material, which determines the chemical reaction environment. The laser-assisted selective functionalization of microcavities can be easily combined with the intensively growing number of laser applications for patterning of molecules and cells, which is useful for the development of novel biological assays.Publicationtype: Journal Article1 - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Access status: without files , High-flexibility combinatorial peptide synthesis with laser-based transfer of monomers in solid matrix material(2016-06-14); ; ; ; Schlageter, MartinLaser writing is used to structure surfaces in many different ways in materials and life sciences. However, combinatorial patterning applications are still limited. Here we present a method for cost-efficient combinatorial synthesis of very-high-density peptide arrays with natural and synthetic monomers. A laser automatically transfers nanometre-thin solid material spots from different donor slides to an acceptor. Each donor bears a thin polymer film, embedding one type of monomer. Coupling occurs in a separate heating step, where the matrix becomes viscous and building blocks diffuse and couple to the acceptor surface. Furthermore, we can consecutively deposit two material layers of activation reagents and amino acids. Subsequent heat-induced mixing facilitates an in situ activation and coupling of the monomers. This allows us to incorporate building blocks with click chemistry compatibility or a large variety of commercially available non-activated, for example, posttranslationally modified building blocks into the array's peptides with >17,000 spots per cm2Publicationtype: Journal Article13