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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
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
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
2014-03-03 08:40:08
date last changed
2019-01-06 04:21:32
@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},
  keyword      = {gels,porous materials,proteins,separation techniques},
  language     = {eng},
  number       = {6},
  pages        = {4383--4391},
  publisher    = {John Wiley & Sons},
  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},
  volume       = {130},
  year         = {2013},
}