Oriented immobilization of peptide ligands on solid supports.
A synthetic procedure was developed for the direct immobilization on preactivated affinity supports of peptidic ligands requiring free alpha-amino groups to recognize their targets properly. The peptidic ligand is assembled by solid-phase peptide synthesis on an octa-branched heptalysine core through a polyglycine spacer, similar to the method developed for the production of multiple antigenic peptides. After deblocking from the resin, peptide is dialysed, lyophylized and used directly for coupling to preactivated supports. Following immobilization, only a limited number of peptide chains are covalently linked to the solid phase, leaving the remainder facing the mobile phase and sufficiently spaced to interact properly. This procedure was applied successfully to the design, synthesis and oriented immobilization of a multimeric tripeptide ligand (Met-Tyr-Phe) for affinity purification of bovine neurophysin.
Fassina
G
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Protein Engineering Unit, Tecnogen ScpA, Milan, Italy.
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000295:instrumentation; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D008969:Molecular Sequence Data; D009481:Neurophysins / Q000302:isolation & purification; D009842:Oligopeptides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D013820:Thermolysin / Q000737:chemistry
D036341:Intercellular Signaling Peptides and Proteins; D008024:Ligands; D009481:Neurophysins; D009842:Oligopeptides; D010455:Peptides; C075229:octameric MYFGGGGG ligand; D013820:Thermolysin