- Setton, L.A. and Ong, S.R. and Trabbic-Carlson, K.A. and Nettles, D.L. and Lim, D.W. and Chilkoti, A., Epitope tagging for tracking elastin-like polypeptides,
Biomaterials (UK), vol. 27 no. 9
pp. 1930 - 5  .
(last updated on 2007/04/10)
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 (His6) epitope tag was incorporated at the N-terminus of an ELP using recombinant DNA techniques to permit tracking without compromising on material biocompatibility. His6-tagged ELPs were successfully detected by Western blot analysis and quantified by ELISAs following digestion with trypsin. The mass of His6 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 His6 tag also facilitated recognition of crosslinked ELPs from background staining of articular cartilage. These results suggest that the His6 epitope tag has the potential to track ELP scaffold loss independently of newly formed tissue mass for evaluating matrix remodeling in vivo. [All rights reserved Elsevier]
biological tissues;biomedical materials;DNA;enzymes;patient treatment;polymer gels;tissue engineering;