Dual Application of Cryogel as Solid Support in Peptide Synthesis and Subsequent Protein-Capture

Gry Jespersen, Anders Laerke Nielsen, Finn Matthiesen, Henrik Sune Andersen, Harald Kirsebom

Research output: Contribution to journalArticlepeer-review

Abstract

The use of a cryogel in a combined application as a solid support for automated synthesis of a peptide ligand followed by affinity chromatography of a target protein is evaluated. The advantage, of synthesizing the ligand directly on the cryogel, is the circumvention of the standard process of synthesizing a peptide on a solid support, followed by cleavage, purification, analysis, and finally immobilization on the cryogel. To demonstrate the application, a peptide affinity ligand is synthesized directly on a cryogel with a yield of 28.4 mol g(-1) dry polymer and purity of 45% of crude product. The affinity capture of an antipeptide antibody reveals a specific binding capacity of 0.86 mg g(-1) dry polymer. To further elucidate the general availability of a peptide ligand to a macromolecular interaction, a trypsin substrate is synthesized on a cryogel. Trypsin cleavage of immobilized substrate is determined to 1.5 mol g(-1) dry polymer. (c) 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 4383-4391, 2013
Original languageEnglish
Pages (from-to)4383-4391
JournalJournal of Applied Polymer Science
Volume130
Issue number6
DOIs
Publication statusPublished - 2013

Subject classification (UKÄ)

  • Industrial Biotechnology

Free keywords

  • gels
  • porous materials
  • proteins
  • separation techniques

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