Dual application of cryogel as solid support in peptide synthesis and subsequent protein-capture

Authors


Correspondence to: H. Kirsebom (E-mail: Harald.Kirsebom@biotek.lu.se)

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 μmol 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 μmol g−1 dry polymer. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 4383–4391, 2013

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