LAUSR

Xenon biosensors are an emerging tool for different molecular imaging approaches. For many applications, their development requires peptide synthesis steps, followed by the selective installation of a xenon host onto the peptide backbone in solution. In this study, three different strategies were attempted for generating entire Xe biosensors on the solid support. Notably, one strategy involving CryA-da was beneficial by directly integrating this host into the growing construct on a low loaded resin via modification of the administered subcomponent equivalents and by prolonging the coupling procedure. Subsequently, installation of additional amino acids or of additional labels onto the growing construct was achieved by a procedure in which an excess amine was administered to the activated CryA-da (acid) anchored onto the resin. Further, the as-generated Xe biosensor was tested for its NMR and MRI capabilities in H2O and compared to the performance of CryA-ma. Xe NMR of the biosensor indicated a clear CEST response and the Xe MR images revealed similar contrast compared to the reference host. These observations suggest that functionalizing CryA-da on both sides with multiple labels did not alter significantly its NMR capabilities. Hereby, we could show the successful and complete synthesis of a CryA-da-based xenon biosensor on the solid support without any notable side reactions and without the necessity of multiple purification steps.