Direct synthesis of peptides on convective interaction media monolithic columns for affinity chromatography.
Solid-phase peptide synthesis was performed on glycidyle methacrylate-co-ethylene dimethacrylate monoliths using Fmoc chemistry. The native epoxy groups were amino-functionalized by reaction with ethylenediamine or ammonia ions. A peptide directed against human blood coagulation factor VIII was synthesized as a model peptide. Amino acid analysis revealed the correct amino acid ratio as present in the sequence. The ligand density of 5 micromol/mL was equal to that achieved with conventional peptide immobilization via epoxy groups. These supports were directly used as peptide affinity chromatography matrixes. The functionality of the CIM monolithic supports was proven by affinity chromatography of factor VIII. The ammonia-functionalized support performed with low hydrophobicity and did not show unspecific adsorption of proteins.
Pflegerl
Karin
K
Institute of Applied Microbiology, University of Agricultural Sciences, Muthgasse 18, A-1190 Vienna, Austria.
Podgornik
Ales
A
Berger
Eva
E
Jungbauer
Alois
A
D000327:Adsorption; D001798:Blood Proteins / Q000302:isolation & purification / Q000493:pharmacokinetics; D002846:Chromatography, Affinity / Q000379:methods; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D005169:Factor VIII / Q000302:isolation & purification* / Q000493:pharmacokinetics; D006801:Humans; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D011108:Polymers / Q000737:chemistry
D001798:Blood Proteins; D010455:Peptides; D011108:Polymers; D005169:Factor VIII