High yield, directed immobilization of a peptide-ligand onto a beaded cellulose support.
Aminopropyl derivatized Perloza beaded cellulose was acylated with alpha-bromoacetic anhydride to give alpha-bromo-acetamidopropyl Perloza. (N-Acetyl)-Cys-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2, the 7 C-terminal amino acids of the decapeptide luteinizing hormone-releasing hormone with a cysteine added to the N-terminus, was synthesized using Fmoc chemistry. The purified peptide (1.35-1.9 eq) was coupled to alpha-bromoacetamidopropyl Perloza in 0.1 M NaHCO3 solution, pH 8.3, for 1-2 hours. The peptide was anchored to the support via a thioether linkage. Analysis of the peptide-Perloza conjugate indicated near-quantitative displacement of support-bound bromine by the peptide. The peptidic affinity matrix was able to bind ovine antibodies to luteinizing hormone-releasing hormone (LHRH). Thioether immobilization offers directed, chemically stable, high-yield anchoring of synthetic peptides onto a chromatographic support. The high reaction efficiency means there is little waste of valuable synthetic peptide.
Englebretsen
D R
DR
Massey University, Palmerston North, New Zealand.
Harding
D R
DR
D000085:Acetates / Q000737:chemistry; D000215:Acylation; D000595:Amino Acid Sequence; D000906:Antibodies / Q000378:metabolism; D001966:Bromine / Q000378:metabolism; D002482:Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D007987:Gonadotropin-Releasing Hormone / Q000737:chemistry / Q000276:immunology; D008024:Ligands; D008863:Microspheres; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D013440:Sulfides
D000085:Acetates; D000906:Antibodies; D008024:Ligands; D010446:Peptide Fragments; D010455:Peptides; D013440:Sulfides; D007987:Gonadotropin-Releasing Hormone; C016274:bromoacetate; D002482:Cellulose; D001966:Bromine