A new combination of protecting groups and links for encoded synthetic libraries suited for consecutive tests on the solid phase and in solution

Abstract

A strategy for high-throughput evaluation of combinatorial compound libraries is reported, which circumvents the necessity to test complex mixtures. The method is based on a new combination of protecting groups, solid-phase linker and tags. The bulk of the library first undergoes a binding assay with the components grafted on beads. A selection of beads carrying strong ligands is stripped from the labelled target and distributed into microvessels. The ligands are cleaved and rinsed into microeluates. Subsequently, a more detailed characterization with a functional assay in solution determines the best performers, which are identified through the peptidic tag left behind on the corresponding mother bead.

Solution‐Phase Combinatorial Libraries: Modulating Cellular Signaling by Targeting Protein–Protein or Protein–DNA Interactions

“…Several advances in the implementation of solid-phase library synthesis paved the way for its widespread acceptance including the spatially arrayed multipin peptide synthesis of Geysen et al, [103,104] the tea-bag method of peptide synthesis introduced by Houghten et al, [105] phage display from Smith et al, [106,107] the spot or disc synthesis developed by Frank et al, [108,109] and especially the (portion-mixing) split-and-mix solid-phase synthesis on beads introduced by Furka et al [110][111][112] and also disclosed by Houghten et al [113] (divide, couple, and recombine), and Lam et al [114][115][116] (split synthesis), and Affymax's light-directed, spatially addressable, immobilized parallel synthesis. [117] Further promoting the widespread acceptance of split-and-mix solid-phase synthesis, several identification techniques were introduced including iterative [110,118,119] and recursive [120] deconvolution, or nucleotide-, [121,222] peptide-, [123][124][125] chemical-, [126][127][128] radiofrequency-, [129,130] color-, [131] and shape-encoded [132] solid-supported libraries. Similarly, advances in microarray synthesis continue to improve large-scale spatially arrayed parallel synthesis on a variety of solid supports.…”

Kombinatorische Flüssigphasensynthese von Bibliotheken: auf der Suche nach Modulatoren für Protein‐Protein‐ und Protein‐DNA‐Wechselwirkungen in der zellulären Signaltransduktion

“…Beispiele für Fortschritte, die der Festphasensynthese von Bibliotheken zu allgemeiner Anerkennung verhalfen, sind die räumlich auflösende Multipin-Peptidsynthese nach Geysen et al, [103,104] die "Teebeutel"-Methode von Houghten et al, [105] das Phagendisplay von Smith et al [106,107] sowie die Spot-oder Disc-Synthese von Frank et al [108,109] Besondere Erwähnung verdienen auch die von Furka et al [110][111][112] eingeführte Split-und-Mix-Festphasensynthese auf Harzkügelchen ("portion-mixing"), an deren Entwicklung auch Houghten et al ("divide, couple, and recombine") [113] sowie Lam et al ("split synthesis") [114][115][116] teilhatten, und die bei Affymax entwickelte lichtgesteuerte, räumlich adressierbare, immobilisierte Parallelsynthese. [117] Neue Analysemethoden trugen zur breiten Annahme der Split-und-Mix-Festphasensynthese bei: die iterative [110,118,119] und die rekursive [120] Dekonvolution, außerdem nucleotid-, [121,122] peptid-, [123][124][125] chemische, [126][127][128] radiofrequenz-, [129,130] farb- [131] und formcodierte [132] festphasengebundene Bibliotheken. Durch Fortschritte bei der Microarray-Herstellung konnte die räumlich auflösende Parallelsynthese im großen Maßstab an verschiedenen Festphasen verbessert werden.…”

Combinatorial Chemistry

“…The tags may be added to provide an encoding sequence whereby the structure of the tag encodes for the building block and the location in the sequence encodes for the library synthesis step (oligonucleotides [9], [101][102][103][104][105] and peptides [106][107][108][109][110] as sequenceable tags, Fig. 12 A).…”