Purification of tryptophan containing synthetic peptides by selective binding of the alpha-amino group to immobilised metal ions.
Immobilised metal ion affinity chromatography (IMAC) based on selective binding via the alpha-amino group to Cu2+ and Ni2+ ions has been used to purify tryptophan containing synthetic peptides. A free alpha-amino group, serving as an affinity handle, is present only in the target peptide when the peptides are synthesised by the solid-phase method and remaining amino groups after each coupling step are blocked by acetylation. A free alpha-amino group is necessary to retain the peptide on the column. The tryptophan residue may contribute to the binding only if the peptide is simultaneously anchored via the alpha-amino group.
Hansen
P
P
Department of Immunology, University of Uppsala, Sweden.
Lindeberg
G
G
D000107:Acetylation; D000588:Amines / Q000737:chemistry*; D000595:Amino Acid Sequence; D002021:Buffers; D002846:Chromatography, Affinity; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D013058:Mass Spectrometry; D008670:Metals / Q000737:chemistry*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000302:isolation & purification*; D014357:Trypsin; D014364:Tryptophan / Q000302:isolation & purification*
D000588:Amines; D002021:Buffers; D008670:Metals; D010455:Peptides; D014364:Tryptophan; D014357:Trypsin