title: Fmoc SPPS using Perloza beaded cellulose.
abstract: Perloza beaded cellulose was functionalised by a cyanoethylation/reduction procedure to give aminopropyl Perloza. Fmoc-amino acids were anchored to aminopropyl Perloza beaded cellulose via the TFA labile 4-oxymethylphenoxyacetyl (HMPA) linker. Using Fmoc-aminoacyl-4-oxymethylphenoxyacetyl-2,4-dichloro-phenyl esters, all 20 amino acids were anchored at substitution levels ranging from 0.37 to 0.65 mmol/g. Fmoc-amino acids were also anchored using the peptide-amide linker 4-[(R,S)-1-[1-(9H-fluoren-9-yl)-methoxycarbonylamino - (2',4'-dimethoxybenzyl]phenoxyacetic acid. The Fmoc-aminoacyl resins were used for SPPS using Fmoc chemistry. SPPS was carried out using either an LKB Biolynx 4175 low-pressure pumped column continuous-flow peptide synthesiser or an ABI 430A automated vortexing batchwise instrument. Comparison of peptides made using each synthesiser showed little difference in quality of the crude peptides. Different Fmoc-amino acid activation methods (DIC/HOBt/DMF, HBTU, DIC/HOBt/DCM) were found to be equally useful with Perloza. Peptides were cleaved using TFA plus scavengers; however, the TFA-swollen resin was not readily separated from the TFA/peptide solution by simple filtration. Therefore alternative cleavage workup procedures were used with Perloza. Peptides were purified by HPLC and characterised by HPLC and amino acid analysis, and in some cases by FAB-MS. Successful syntheses ranged from 5 to 34 amino acids in length. Some of the peptides were also synthesized using a polystyrene support and standardised (ABI Fastmoc) SPPS protocols. The crude cleaved peptides from each synthesis were compared by HPLC analysis. The overall aim of our work with Perloza is synthesis of resin-bound peptide ligands for affinity chromatography and antibody generation.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Separation Science Unit, Massey University, Palmerston North, New Zealand.
authors.forename: D R
authors.initials: DR
authors.lastname: Englebretsen
authors.forename: D R
authors.initials: DR
authors.lastname: Harding
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids; D000906:Antibodies / Q000737:chemistry; D002482:Cellulose; D002846:Chromatography, Affinity / Q000379:methods; D005449:Fluorenes; D006492:Hempa; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis*; D012116:Resins, Plant
chemical_list: D000596:Amino Acids; D000906:Antibodies; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D010455:Peptides; D012116:Resins, Plant; D002482:Cellulose; D006492:Hempa