Epitope tagging for tracking elastin-like polypeptides

Ong, Shin R.; Trabbic‐Carlson, Kimberly; Nettles, Dana L.; Lim, Dong Woo; Chilkoti, Ashutosh; Setton, Lori A.

doi:10.1016/j.biomaterials.2005.10.018

Cited by 51 publications

(43 citation statements)

References 23 publications

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“…Because ELPs are genetically encoded, their sequence and molecular weight can be precisely controlled, and modulation of these parameters allows the ELP transition behavior to be precisely tuned, which in turn controls its in vivo half-life [27]. Importantly, protein-ELP fusions have been shown to maintain temperature responsive behavior [29] and the ELPs themselves are biodegradable [30] and non-immunogenic [31]. In addition, ELP fusions can be purified to very high purity without the need for chromatography by a batch purification process –termed inverse transition cycling (ITC)– that takes advantage of their reversible soluble-insoluble phase transition behavior [32].…”

Section: Resultsmentioning

confidence: 99%

A depot-forming glucagon-like peptide-1 fusion protein reduces blood glucose for five days with a single injection

Amiram

Luginbuhl

et al. 2013

Journal of Controlled Release

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Peptide drugs are an exciting class of pharmaceuticals for the treatment of a variety of diseases; however, their short half-life dictates multiple and frequent injections causing undesirable side-effects. Herein, we describe a novel peptide delivery system that seeks to combine the attractive features of prolonged circulation time with a prolonged release formulation. This system consists of glucagon-like peptide-1, a type-2 diabetes drug fused to a thermally responsive, elastin-like-polypeptide (ELP) that undergoes a soluble-insoluble phase transition between room temperature and body temperature, thereby forming an injectable depot. We synthesized a set of GLP-1-ELP fusions and verified their proteolytic stability and potency in vitro. Significantly, a single injection of depot forming GLP-1-ELP fusions reduced blood glucose levels in mice for up to 5 days, 120 times longer than an injection of the native peptide. These findings demonstrate the unique advantages of using ELPs to release peptide-ELP fusions from a depot combined with enhanced systemic circulation to create a tunable peptide delivery system.

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Section: Resultsmentioning

confidence: 99%

A depot-forming glucagon-like peptide-1 fusion protein reduces blood glucose for five days with a single injection

Amiram

Luginbuhl

et al. 2013

Journal of Controlled Release

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“…The ELP hydrogel was largely absent from the defect by 3 months, however, likely related to proteolytically mediated degradation of the ELP. 42 This may explain why repair in control defects outpaced ELP-filled defects at the 6-month time point. Because ELPs are genetically engineered, the degradation, crosslink density, transition temperature, and other properties of these gels are parameters that may be altered in a well-controlled manner to achieve a slower degradation rate.…”

Section: Discussionmentioning

confidence: 99%

In Situ Crosslinking Elastin-Like Polypeptide Gels for Application to Articular Cartilage Repair in a Goat Osteochondral Defect Model

Nettles

Kitaoka

Hanson

et al. 2008

Tissue Engineering Part A

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The objective of this study was to evaluate an injectable, in situ crosslinkable elastin-like polypeptide (ELP) gel for application to cartilage matrix repair in critically sized defects in goat knees. One cylindrical, osteochondral defect in each of seven animals was filled with an aqueous solution of ELP and a biocompatible, chemical crosslinker, while the contralateral defect remained unfilled and served as an internal control. Joints were sacrificed at 3 (n = 3) or 6 (n = 4) months for MRI, histological, and gross evaluation of features of biomaterial performance, including integration, cellular infiltration, surrounding matrix quality, and new matrix in the defect. At 3 months, ELP-filled defects scored significantly higher for integration by histological and gross grading compared to unfilled defects. ELP did not impede cell infiltration but appeared to be partly degraded. At 6 months, new matrix in unfilled defects outpaced that in ELP-filled defects and scored significantly better for MRI evidence of adverse changes, as well as integration and proteoglycan-containing matrix via gross and histological grading. The ELP-crosslinker solution was easily delivered and formed stable, well-integrated gels that supported cell infiltration and matrix synthesis; however, rapid degradation suggests that ELP formulation modifications should be optimized for longer-term benefits in cartilage repair applications.

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“…However, it was reported by Lim et al that an aqueous, biocompatible Mannich type condensation reaction [265][266][267] can be used to crosslink lysine-containing ELPs over a wide range in pH (2-13), and it produces only water as a byproduct [260,268]. In this reaction, an organophosphorous crosslinker, β-[tris(hydroxymethyl)phosphino]propionic acid (THPP), reacts with the amines of the lysine residues in the ELP to create trifunctional intraor intermolecular crosslinks, as shown in Figure 21.…”

Section: Chemically Crosslinked Elastin-based Hydrogelsmentioning

confidence: 99%

Peptide-based biopolymers in biomedicine and biotechnology

Chow

Nunalee

Lim

et al. 2008

Materials Science and Engineering: R: Reports

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Peptides are emerging as a new class of biomaterials due to their unique chemical, physical, and biological properties. The development of peptide-based biomaterials is driven by the convergence of protein engineering and macromolecular self-assembly. This review covers the basic principles, applications, and prospects of peptide-based biomaterials. We focus on both chemically synthesized and genetically encoded peptides, including poly-amino acids, elastin-like polypeptides, silk-like polymers and other biopolymers based on repetitive peptide motifs. Applications of these engineered biomolecules in protein purification, controlled drug delivery, tissue engineering, and biosurface engineering are discussed.

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Epitope tagging for tracking elastin-like polypeptides

Cited by 51 publications

References 23 publications

A depot-forming glucagon-like peptide-1 fusion protein reduces blood glucose for five days with a single injection

A depot-forming glucagon-like peptide-1 fusion protein reduces blood glucose for five days with a single injection

In Situ Crosslinking Elastin-Like Polypeptide Gels for Application to Articular Cartilage Repair in a Goat Osteochondral Defect Model

Peptide-based biopolymers in biomedicine and biotechnology

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