Automated Laser‐Transfer Synthesis of High‐Density Microarrays for Infectious Disease Screening

Abstract

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−2. 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.

Laser-Induced Transfer of Functional Materials

“…Peptide microarrays with thousands of distinct sequences can be synthesized using the LIFT system. A LIFT-based microarray synthesizer was developed to produce up to 20 residue peptides with a density of 10,000 spots per cm 2 [ 45 ]. These produced peptide microarrays were used to study the IgG response in a survivor of Ebola virus sickness.…”

Application of Digital Methods in Polymer Science and Engineering

“…A smaller version of this approach is the integration of a robotic arm in the synthesis setup for reactions in a microarray format carried out in parallel. [ 32 ] An increasing possibility of online characterization, as is already known for organic chemistry or flow chemistry, [ 33 ] would also be desirable for robot‐based chemistry in order to further eliminate the bottleneck of characterization. The approach of linking flow chemistry with robot chemistry definitely represents a convincing approach.…”

Screening for Primordial RNA–Peptide Interactions Using High-Density Peptide Arrays

“…However, the statistically derived amino acid/nucleotide interaction preferences can be strongly biased toward the stable ribonucleoprotein complexes as a product of the modern ribosomal translation. In this work, we demonstrate a bottom-up approach to studying primordial RNA/peptide interactions with fully combinatorial high-density peptide arrays [ 25 , 26 , 27 , 28 ] to evaluate as many potential interactions as possible.…”