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Epitope tagging for tracking elastin-like polypeptides

Paper ID Volume ID Publish Year Pages File Format Full-Text
10785 702 2006 6 PDF Available
Title
Epitope tagging for tracking elastin-like polypeptides
Abstract

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.

Keywords
Elastin-like polypeptide; Oligohistidine tag; ELISA; Degradation; Crosslinking; Cartilage repairELP, elastin-like polypeptide; His6, Oligohistidine THPP; β-[Tris (hydroxymethyl) phosphino] propionic acid
First Page Preview
Epitope tagging for tracking elastin-like polypeptides
Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 27, Issue 9, March 2006, Pages 1930–1935
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering