A chemoselective method for site-specific immobilization of peptides via aminooxy group.
Site-specific modification of peptides and proteins is an important area of basic research for preparation of well-defined biosensors and probes. The unique properties of aminooxy group present an opportunity for chemoselective site-specific immobilization of peptides to prepare well-defined biosensors. We have prepared FLAG peptide derivatives containing L-epsilon-aminooxylysine (L-epsilon-AOLys, 1a) and L-lysine units in their sequence at the C- and N-terminals via solid-phase synthesis. Site-specific modification of peptides through aminooxy group was demonstrated in the preparation of biosensors and selective conjugation in the preparation of biotinylated probes. Effect of the incorporation of L-epsilon-AOLys (1a) into the peptide sequence and its subsequent labeling on the FLAG epitopic character was measured using a surface plasmon resonance detector. It was found that incorporation of L-epsilon-AOLys (1a) into the FLAG peptide and site-specific immobilization through aminooxy group preserved the integrity of FLAG epitope.
Adamczyk
M
M
Department of Chemistry, 9NM, Building AP20, Diagnostics Division, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, Illinois 60064-6016, USA. maciej.adamczyk@abbott.com
Gebler
J C
JC
Reddy
R E
RE
Yu
Z
Z
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D015374:Biosensing Techniques; D000939:Epitopes; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry; D016297:Mutagenesis, Site-Directed; D009842:Oligopeptides; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D020349:Surface Plasmon Resonance
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009842:Oligopeptides; D010455:Peptides; C085706:FLAG peptide; D008239:Lysine