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

Jespersen, Gry LU ; Nielsen, Anders Laerke ; Matthiesen, Finn ; Andersen, Henrik Sune and Kirsebom, Harald LU (2013) In Journal of Applied Polymer Science 130(6). p.4383-4391
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... (More)
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 (Less)
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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
gels, porous materials, proteins, separation techniques
in
Journal of Applied Polymer Science
volume
130
issue
6
pages
4383 - 4391
publisher
John Wiley & Sons Inc.
external identifiers
  • wos:000329153300063
  • scopus:84885018912
ISSN
1097-4628
DOI
10.1002/app.39727
language
English
LU publication?
yes
id
6ea395b4-c0a6-4d45-aa7e-9e3b1e4c139b (old id 4327132)
date added to LUP
2016-04-01 10:33:32
date last changed
2022-01-26 00:22:40
@article{6ea395b4-c0a6-4d45-aa7e-9e3b1e4c139b,
  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}},
  author       = {{Jespersen, Gry and Nielsen, Anders Laerke and Matthiesen, Finn and Andersen, Henrik Sune and Kirsebom, Harald}},
  issn         = {{1097-4628}},
  keywords     = {{gels; porous materials; proteins; separation techniques}},
  language     = {{eng}},
  number       = {{6}},
  pages        = {{4383--4391}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{Journal of Applied Polymer Science}},
  title        = {{Dual Application of Cryogel as Solid Support in Peptide Synthesis and Subsequent Protein-Capture}},
  url          = {{http://dx.doi.org/10.1002/app.39727}},
  doi          = {{10.1002/app.39727}},
  volume       = {{130}},
  year         = {{2013}},
}