SUBPOL: a novel SUcrose-Based Polymer support for solid-phase peptide synthesis and affinity chromatography applications.
A novel SUcrose-Based Polymer support (SUBPOL) with tailored morphology suitable for the use in solid-phase peptide synthesis (SPPS) is described, and its application as a hydrophilic affinity matrix for the specific removal of fibrinogen from human plasma is demonstrated. After suspension polymerization of partly methacrylated 2,1':4,6-di-O-isopropylidene sucrose and subsequent removal of the protecting groups, hydrophilic spherical polymer beads were obtained. The morphology of the resulting resin was controlled by variation of the porogen as well as the average degree of substitution with respect to the methacryloyl groups of the monomer mixture. After introduction of amino groups for a permanent attachment of immobilized peptide ligands, prevention of unintended esterification during SPPS was achieved by silylation of remaining hydroxy groups. Alternatively, a Rink amide linker was introduced prior to SPPS to allow cleavage and subsequent analysis of the peptide assembled on the SUBPOL resins. Two hexapeptides of sequence kwiivw and hffflw, consisting of d-amino acids, as well as a 19-mer peptide corresponding to the sequence GSGVRGDFGSLAPRVARQL of the VP1 protein from the foot-and-mouse disease virus (FMDV) were successfully prepared both manually or in a semi-automated process on SUBPOL resins according to the Fmoc/tBu strategy. Yields and purities were comparable to peptides prepared on commercially available polystyrene resins. A specific affinity adsorbent containing the fibrinogen-binding pentapeptide GPRPK was prepared by SPPS on SUBPOL resins of different morphology, and the strong impact of the affinity matrix on adsorption performance was demonstrated.
Poschalko
Alexander
A
Vienna University of Technology, Institute of Applied Synthetic Chemistry, Division of Macromolecular Chemistry, Getreidemarkt 9/163/MC, 1060 Vienna, Austria. Alexander.Poschalko@roche.com
Rohr
Thomas
T
Gruber
Heinrich
H
Bianco
Alberto
A
Guichard
Gilles
G
Briand
Jean-Paul
JP
Weber
Viktoria
V
Falkenhagen
Dieter
D
D000595:Amino Acid Sequence; D036022:Capsid Proteins / Q000138:chemical synthesis; D002846:Chromatography, Affinity / Q000379:methods*; D005340:Fibrinogen / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis; D010455:Peptides / Q000138:chemical synthesis*; D011109:Polymethacrylic Acids / Q000138:chemical synthesis / Q000737:chemistry*; D013395:Sucrose / Q000031:analogs & derivatives* / Q000737:chemistry
D036022:Capsid Proteins; D010446:Peptide Fragments; D010455:Peptides; D011109:Polymethacrylic Acids; C442501:VP1 protein, Foot-and-mouth disease virus; C030613:polymethacrylic acid; D013395:Sucrose; D005340:Fibrinogen