A novel combinatorial approach to high-density peptide arrays.
Combinatorial synthesis of peptides on solid supports (1), either as spots on cellulose membranes (2) or with split-pool-libraries on polymer beads (3), substantially forwarded research in the field of peptide-protein interactions. Admittedly, these concepts have specific limitations, on one hand the number of synthesizable peptide sequences per area, on the other hand elaborate decoding/encoding strategies, false-positive results and sequence limitations. We recently established a method to produce high-density peptide arrays on microelectronic chips (4). Solid amino acid microparticles were charged by friction and transferred to defined pixel electrodes onto the chip's surface, where they couple to a functional polymer coating simply upon melting (Fig. 16.1 A-D,F). By applying standard Fmoc chemistry according to Merrifield, peptide array densities of up to 40,000 spots per square centimetre were achieved (Fig. 16.1G). The term "Merrifield synthesis" describes the consecutive linear coupling and deprotecting of L-amino acids modified with base-labile fluorenylmethoxy (Fmoc) groups at the N-terminus and different acid-sensitive protecting groups at their side chains. Removing side chain protecting groups takes place only once at the very end of each synthesis and generates the natural peptide sequence thereby.
Beyer
Mario
M
Department of Chip-Based Peptide Libraries, German Cancer Research Center, Heidelberg, Germany.
Block
Ines
I
König
Kai
K
Nesterov
Alexander
A
Fernandez
Simon
S
Felgenhauer
Thomas
T
Schirwitz
Christopher
C
Leibe
Klaus
K
Bischoff
Ralf F
RF
Breitling
Frank
F
Stadler
Volker
V
D000818:Animals; D001798:Blood Proteins / Q000494:pharmacology; D020650:Combinatorial Chemistry Techniques / Q000295:instrumentation / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008689:Methacrylates / Q000494:pharmacology; D008954:Models, Biological; D019151:Peptide Library; D011092:Polyethylene Glycols / Q000494:pharmacology; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods; D013499:Surface Properties
D001798:Blood Proteins; D008689:Methacrylates; D019151:Peptide Library; C524499:polyethylene glycol methacrylate; C028035:stable plasma protein solution; D011092:Polyethylene Glycols