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Epitope tagging for tracking elastin-like polypeptides.

Publication ,  Journal Article
Ong, SR; Trabbic-Carlson, KA; Nettles, DL; Lim, DW; Chilkoti, A; Setton, LA
Published in: Biomaterials
March 2006

Elastin-like polypeptides (ELPs) are a class of biocompatible, non-immunogenic and crosslinkable biomaterials that offer promise for use as an injectable scaffold for cartilage repair. In this study, an oligohistidine (His(6)) epitope tag was incorporated at the N-terminus of an ELP using recombinant DNA techniques to permit tracking without compromising on material biocompatibility. His(6)-tagged ELPs were successfully detected by Western blot analysis and quantified by ELISAs following digestion with trypsin. The mass of His(6) tagged ELP fragments freed from a crosslinked ELP hydrogel after digestion with trypsin correlated highly with hydrogel weight loss, providing evidence of the tag's capability to enable tracking of enzymatic degradation of the ELP hydrogel. The His(6) tag also facilitated recognition of crosslinked ELPs from background staining of articular cartilage. These results suggest that the His(6) epitope tag has the potential to track ELP scaffold loss independently of newly formed tissue mass for evaluating matrix remodeling in vivo.

Duke Scholars

Published In

Biomaterials

DOI

EISSN

1878-5905

ISSN

0142-9612

Publication Date

March 2006

Volume

27

Issue

9

Start / End Page

1930 / 1935

Related Subject Headings

  • Trypsin
  • Swine
  • Protein Engineering
  • Peptides
  • Molecular Sequence Data
  • Hydrogels
  • Epitopes
  • Cartilage, Articular
  • Biomedical Engineering
  • Biodegradation, Environmental
 

Citation

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Ong, S. R., Trabbic-Carlson, K. A., Nettles, D. L., Lim, D. W., Chilkoti, A., & Setton, L. A. (2006). Epitope tagging for tracking elastin-like polypeptides. Biomaterials, 27(9), 1930–1935. https://doi.org/10.1016/j.biomaterials.2005.10.018
Ong, Shin R., Kimberly A. Trabbic-Carlson, Dana L. Nettles, Dong W. Lim, Ashutosh Chilkoti, and Lori A. Setton. “Epitope tagging for tracking elastin-like polypeptides.Biomaterials 27, no. 9 (March 2006): 1930–35. https://doi.org/10.1016/j.biomaterials.2005.10.018.
Ong SR, Trabbic-Carlson KA, Nettles DL, Lim DW, Chilkoti A, Setton LA. Epitope tagging for tracking elastin-like polypeptides. Biomaterials. 2006 Mar;27(9):1930–5.
Ong, Shin R., et al. “Epitope tagging for tracking elastin-like polypeptides.Biomaterials, vol. 27, no. 9, Mar. 2006, pp. 1930–35. Epmc, doi:10.1016/j.biomaterials.2005.10.018.
Ong SR, Trabbic-Carlson KA, Nettles DL, Lim DW, Chilkoti A, Setton LA. Epitope tagging for tracking elastin-like polypeptides. Biomaterials. 2006 Mar;27(9):1930–1935.
Journal cover image

Published In

Biomaterials

DOI

EISSN

1878-5905

ISSN

0142-9612

Publication Date

March 2006

Volume

27

Issue

9

Start / End Page

1930 / 1935

Related Subject Headings

  • Trypsin
  • Swine
  • Protein Engineering
  • Peptides
  • Molecular Sequence Data
  • Hydrogels
  • Epitopes
  • Cartilage, Articular
  • Biomedical Engineering
  • Biodegradation, Environmental