Combinatorial peptide synthesis on a microchip.
Microchips are used in the combinatorial synthesis of peptide arrays by means of amino acid microparticle deposition. The surface of custom-built microchips can be equipped with an amino-modified poly(ethylene glycol)methacrylate (PEGMA) graft polymer coating, which permits high loading of functional groups and resists nonspecific protein adsorption. Specific microparticles that are addressed to the polymer-coated microchip surface in a well defined pattern release preactivated amino acids upon melting, and thus allow combinatorial synthesis of high-complexity peptide arrays directly on the chip surface. Currently, arrays with densities of up to 40,000 peptide spots/cm(2) can be generated in this way, with a minimum of coupling cycles required for full combinatorial synthesis. Without using any additional blocking agent, specific peptide recognition has been verified by background-free immunostaining on the chip-based array. This unit describes microchip surface modification, combinatorial peptide array synthesis on the chip, and a typical immunoassay employing the resulting high-density peptide arrays.
Schirwitz
Christopher
C
German Cancer Research Center, Heidelberg, Germany.
Block
Ines
I
German Cancer Research Center, Heidelberg, Germany.
König
Kai
K
German Cancer Research Center, Heidelberg, Germany.|University of Heidelberg, Kirchhoff Institute for Physics, Heidelberg, Germany.
Nesterov
Alexander
A
German Cancer Research Center, Heidelberg, Germany.|University of Heidelberg, Kirchhoff Institute for Physics, Heidelberg, Germany.
Fernandez
Simon
S
German Cancer Research Center, Heidelberg, Germany.
Felgenhauer
Thomas
T
German Cancer Research Center, Heidelberg, Germany.
Leibe
Klaus
K
German Cancer Research Center, Heidelberg, Germany.
Torralba
Gloria
G
University of Heidelberg, Kirchhoff Institute for Physics, Heidelberg, Germany.
Hausmann
Michael
M
University of Heidelberg, Kirchhoff Institute for Physics, Heidelberg, Germany.
Lindenstruth
Volker
V
University of Heidelberg, Kirchhoff Institute for Physics, Heidelberg, Germany.
Stadler
Volker
V
German Cancer Research Center, Heidelberg, Germany.
Breitling
Frank
F
German Cancer Research Center, Heidelberg, Germany.
Bischoff
F Ralf
FR
German Cancer Research Center, Heidelberg, Germany.
D000327:Adsorption; D000596:Amino Acids / Q000737:chemistry; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D020650:Combinatorial Chemistry Techniques; D008689:Methacrylates / Q000737:chemistry; D010316:Particle Size; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D011506:Proteins / Q000737:chemistry; D013499:Surface Properties
D000596:Amino Acids; D020099:Coated Materials, Biocompatible; D008689:Methacrylates; D010455:Peptides; D011108:Polymers; D011506:Proteins; C524499:polyethylene glycol methacrylate; D011092:Polyethylene Glycols
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