Automated laser-transfer synthesis of high-density microarrays for infectious disease screening
| Autor: | Grigori Paris, Jasmin Heidepriem, Alexandra Tsouka, Yuxin Liu, Daniela S. Mattes, Sandra Pinzón Martín, Pietro Dallabernardina, Marco Mende, Celina Lindner, Robert Wawrzinek, Christoph Rademacher, Peter H. Seeberger, Frank Breitling, Frank Ralf Bischoff, Timo Wolf, Felix F. Loeffler |
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| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
Mechanical Engineering
Lasers Reproducibility of Results 500 Naturwissenschaften und Mathematik::570 Biowissenschaften Biologie::570 Biowissenschaften Biologie Hemorrhagic Fever Ebola Schiff base fluorophores high‐throughput Communicable Diseases solid phase synthesis laser‐induced forward transfer 540 Chemie und zugeordnete Wissenschaften Mechanics of Materials laser-induced forward transfer peptides Humans General Materials Science ddc:620 Engineering & allied operations high-throughput |
| Zdroj: | Advanced Materials Advanced Materials, 34 (23), Art.Nr. 2200359 |
| ISSN: | 0935-9648 1521-4095 |
| Popis: | 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 limited LIFT applications to simple proof-of-concept syntheses of fewer than 100 compounds. Here, we report an automated synthesis instrument that combines laser transfer and robotics for parallel synthesis in a microarray format with up to 10000 individual reactions/cm2. 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 were synthesized to study the antibody response of an Ebola virus infection survivor. We identified known and unknown epitopes that serve now as a basis for Ebola diagnostic development. 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. |
| Databáze: | OpenAIRE |
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