High-flexibility combinatorial peptide synthesis with laser-based transfer of monomers in solid matrix material

Abstract

Laser 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 cm2.

A strategy to identify linker-based modules for the allosteric regulation of antibody-antigen binding affinities of different scFvs

“…Moreover, the selection and screening of new calmodulin-binding peptides in a library approach may lead to improved combinations of permuted CaM variants and corresponding binding peptides. This could be achieved by creation of peptide libraries and screening for the desired characteristics by phage display 46 or very-high-density peptide arrays 48 . Another approach to enhance the modulation may be the replacement of calcium with other ions known to bind calmodulin and induce activation of binding partners 49,50 .…”

A Simple Platform for the Rapid Development of Antimicrobials

“…Some of the features demonstrated here, for example the rapid synthesis of a large number of synbody candidates, could also be used in standard approaches to therapeutic development. Alternatively, parallel advances in high density peptide microarray synthesis [33][34][35][36][37][38] and rapid peptide synthesis 39 , could dramatically increase the diversity of the peptide screening and decrease time to synthesize the resulting hits. These emerging technologies could eliminate the need for library pre-synthesis and yet maintain the speed of the discovery platform, increasing the potential for this approach.…”

Screening for Selective Protein Inhibitors by Using the IANUS Peptide Array

“…To screen all possible peptide combinations of both proteins (overlapping 12‐mer peptides that fully scan through each protein sequence with a frame shift of two amino acids), an excessive number of peptide pairs would have been needed (89 890 peptide pairs). The parallel synthesis of such large numbers of peptides cannot be easily achieved, even with modern lithographic methods . Instead, the SGPSPRIEITPSH peptide, which contained the SPRIEIT sequence from the regulatory domain of NFAT, was used to screen for CaN‐interacting epitopes.…”