Multifunctional Materials through Modular Protein Engineering
ABSTRACT The diversity of potential applications for protein-engineered materials has undergone profound recent expansion through a rapid increase in the library of domains that have been utilized in these materials. Historically, protein-engineered biomaterials have been generated from a handful of peptides that were selected and exploited for their naturally evolved functionalities. In recent years, the scope of the field has drastically expanded to include peptide domains that were designed through computational modeling, identified through high-throughput screening, or repurposed from wild type domains to perform functions distinct from their primary native applications. The strategy of exploiting a diverse library of peptide domains to design modular block copolymers enables the synthesis of multifunctional protein-engineered materials with a range of customizable properties and activities. As the diversity of peptide domains utilized in modular protein engineering continues to expand, a tremendous and ever-growing combinatorial expanse of material functionalities will result.
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ABSTRACT: Hydrogels have strong application prospects for drug delivery, tissue engineering and cell therapy because of their excellent biocompatibility and abundant availability as scaffolds for drugs and cells. In this study, we created hybrid hydrogels based on a genetically modified tax interactive protein-1 (TIP1) by introducing two or four cysteine residues in the primary structure of TIP1. The introduced cysteine residues were crosslinked with a four-armed poly (ethylene glycol) having their arm ends capped with maleimide residues (4-armed-PEG-Mal) to form hydrogels. In one form of the genetically modification, we incorporated a peptide sequence 'GRGDSP' to introduce bioactivity to the protein, and the resultant hydrogel could provide an excellent environment for a three dimensional cell culture of AD293 cells. The AD293 cells continued to divide and displayed a polyhedron or spindle-shape during the 3-day culture period. Besides, AD293 cells could be easily separated from the cell-gel constructs for future large-scale culture after being cultured for 3 days and treating hydrogel with trypsinase. This work significantly expands the toolbox of recombinant proteins for hydrogel formation, and we believe that our hydrogel will be of considerable interest to those working in cell therapy and controlled drug delivery.PLoS ONE 09/2014; 9(9):e107949. DOI:10.1371/journal.pone.0107949 · 3.53 Impact Factor
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ABSTRACT: The density of integrin-binding ligands in an extracellular matrix (ECM) is known to regulate cell migration speed by imposing a balance of traction forces between the leading and trailing edges of the cell, but the effect of cell-adhesive ligands on neurite chemoattraction is not well understood. A platform is presented here that combines gradient-generating microfluidic devices with 3D protein-engineered hydrogels to study the effect of RGD ligand density on neurite pathfinding from chick dorsal root ganglia-derived spheroids. Spheroids are encapsulated in elastin-like polypeptide (ELP) hydrogels presenting either 3.2 or 1.6 mM RGD ligands and exposed to a microfluidic gradient of nerve growth factor (NGF). While the higher ligand density matrix enhanced neurite initiation and persistence of neurite outgrowth, the lower ligand density matrix significantly improved neurite pathfinding and increased the frequency of growth cone turning up the NGF gradient. The apparent trade-off between neurite extension and neurite guidance is reminiscent of the well-known trade-off between adhesive forces at the leading and trailing edges of a migrating cell, implying that a similar matrix-mediated balance of forces regulates neurite elongation and growth cone turning. These results have implications in the design of engineered materials for in vitro models of neural tissue and in vivo nerve guidance channels.Small 10/2014; 11(6). DOI:10.1002/smll.201401574 · 7.51 Impact Factor
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ABSTRACT: Nature, through evolution over millions of years, has perfected materials with amazing characteristics and awe-inspiring functionalities that exceed the performance of man-made synthetic materials. One such remarkable material is native resilin – an extracellular skeletal protein that plays a major role in the jumping, flying, and sound production mechanisms in many insects. It is one of the most resilient (energy efficient) elastomeric biomaterials known with a resilience of [similar]97% and a fatigue life in excess of 300 million cycles. Recently, resilin-like polypeptides (RLPs) with exquisite control over the amino acid sequence (comprising repeat resilin motifs) and tuneable biological properties and/or functions have been generated by genetic engineering and cloning techniques. RLPs have been the subject of intensive investigation over a decade and are now recognized to be multi-functional and multi-stimuli responsive; including temperature (exhibiting both an upper and a lower critical solution temperature), pH, moisture, ion and photo-responsive with tuneable photo-physical properties. Such unusual multi-stimuli responsiveness has scarcely been offered and reported for either synthetic or natural biopolymers. Furthermore, the directed molecular self-assembly property of RLPs also exhibits promise as efficient templates for the synthesis and stabilization of metal nanoparticles. These developments and observations reveal the opportunities and new challenges for RLPs as novel materials for nanotechnology, nanobiotechnology and therapeutic applications. In this review, we discuss and highlight the design and synthesis of different RLPs, their unique molecular architecture, advanced responsive behaviour, and functionality of hydrogels, solid–liquid interfaces, nanoparticles and nanobioconjugates derived from RLPs.06/2014; 2(36):5936-5947. DOI:10.1039/C4TB00726C