Combinatorial libraries of peptide dendrimers: design, synthesis, on-bead high-throughput screening, bead decoding and characterization

Abstract

Dendrimers are branched synthetic macromolecules. This protocol describes the synthesis (1-2 weeks), functional screening (1.5 d) and decoding (2 d) of 'one-bead-one-compound' combinatorial libraries of dendrimers assembled from amino-acid building blocks by 'split-and-mix' solid phase peptide synthesis. The method resembles that for synthesizing linear peptides, except that a branching diamino acid is used at every third position to obtain the dendritic structure. Structural diversification by splitting is restricted to four amino acids per variable position, yielding libraries of approximately 60,000 sequences. In such libraries, the sequence of a dendrimer can be deduced uniquely from an amino-acid analysis of the solid support bead. This analysis is more reliable, faster and far less costly than Edman sequencing such that decoding multiple beads is affordable. The method is exemplified for the identification of catalytic peptide dendrimers catalyzing the hydrolysis of acyloxypyrene-trisulfonates with substrate binding (K(M) = 10-300 microM) and rate accelerations up to k(cat)/k(uncat) = 10(4) in aqueous buffer.

Peptide-Based Polymer Therapeutics

“…One example for this application is the work done by Reimond et al [163][164][165][166]. They have identified a set of peptide-based dendrimers, firstly made from His-Ser and diaminopropionic acid (Dap) as branching units.…”

The proteome buccaneers: how to unearth your treasure chest via combinatorial peptide ligand libraries

“…With one proviso, a library composed by 16 different amino acids would contain 16.7 million diversomers. For all of them to be represented, one would need to operate with approximately 4.5 ml of beads of 65-µm diameter or, according to Maillard et al [68] approximately 11 ml. In most of our work, we have used 1 ml of settled beads, comprising, thus ca.…”

Other Applications of Combinatorial Peptide Libraries

“…To the question of peptide bead screening, Maillard et al (2009) have provided additional insights into peptide libraries. Table 7.2 summarizes the advantages and disadvantages of bead identification.…”