Elastin‐like recombinamers: Biosynthetic strategies and biotechnological applications

The past few decades have witnessed the development of novel naturally inspired biomimetic materials, such as polysaccharides and proteins. Likewise, the seemingly exponential evolution of genetic‐engineering techniques and modern biotechnology has led to the emergence of advanced protein‐based materials with multifunctional properties. This approach allows extraordinary control over the architecture of the polymer, and therefore, monodispersity, controlled physicochemical properties, and high sequence complexity that would otherwise be impossible to attain. Elastin‐like recombinamers (ELRs) are emerging as some of the most prolific of these protein‐based biopolymers. Indeed, their inherent properties, such as biocompatibility, smart nature, and mechanical qualities, make these recombinant polymers suitable for use in numerous biomedical and nanotechnology applications, such as tissue engineering, “smart” nanodevices, drug delivery, and protein purification. Herein, we present recent progress in the biotechnological applications of ELRs and the most important genetic engineering‐based strategies used in their biosynthesis.

[1]  J. Rodríguez‐Cabello,et al.  “Recombinamers” as advanced materials for the post-oil age , 2009 .

[2]  R. Herrero-Vanrell,et al.  Self-assembled particles of an elastin-like polymer as vehicles for controlled drug release. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[3]  José Carlos Rodríguez-Cabello,et al.  Recombinamers: combining molecular complexity with diverse bioactivities for advanced biomedical and biotechnological applications. , 2011, Advances in biochemical engineering/biotechnology.

[4]  Ashutosh Chilkoti,et al.  Applications of elastin-like polypeptides in tissue engineering. , 2010, Advanced drug delivery reviews.

[5]  Wilfred Chen,et al.  Cadmium removal from contaminated soil by tunable biopolymers. , 2004, Environmental Science and Technology.

[6]  G. Bidwell,et al.  Targeting a c-Myc inhibitory polypeptide to specific intracellular compartments using cell penetrating peptides. , 2009, Journal of controlled release : official journal of the Controlled Release Society.

[7]  Joseph Cappello,et al.  Genetic Production of Synthetic Protein Polymers , 1992 .

[8]  E. Angov,et al.  Adjustment of codon usage frequencies by codon harmonization improves protein expression and folding. , 2011, Methods in molecular biology.

[9]  S. Opal Endotoxins and other sepsis triggers. , 2010, Contributions to Nephrology.

[10]  E. Kobatake,et al.  Design and construction of glutamine binding proteins with a self-adhering capability to unmodified hydrophobic surfaces as reagentless fluorescence sensing devices. , 2003, Journal of the American Chemical Society.

[11]  Maido Remm,et al.  Preferred and avoided codon pairs in three domains of life , 2008, BMC Genomics.

[12]  Wilfred Chen,et al.  Genetic engineering of self-assembled protein hydrogel based on elastin-like sequences with metal binding functionality. , 2007, Biomacromolecules.

[13]  Rui R. Costa,et al.  Stimuli‐Responsive Thin Coatings Using Elastin‐Like Polymers for Biomedical Applications , 2009 .

[14]  T. M. Parker,et al.  Biocompatibility of the Bioelastic Materials, Poly(GVGVP) and Its γ-Irradiation Cross-Linked Matrix: Summary of Generic Biological Test Results , 1991 .

[15]  J. Rodríguez‐Cabello,et al.  Effect of NaCl on the exothermic and endothermic components of the inverse temperature transition of a model elastin-like polymer. , 2007, Biomacromolecules.

[16]  G. Bidwell,et al.  Cell penetrating elastin-like polypeptides for therapeutic peptide delivery. , 2010, Advanced drug delivery reviews.

[17]  Ashutosh Chilkoti,et al.  Purification of recombinant proteins by fusion with thermally-responsive polypeptides , 1999, Nature Biotechnology.

[18]  Stefan Rose-John,et al.  Elastin-like polypeptides revolutionize recombinant protein expression and their biomedical application. , 2010, Trends in biotechnology.

[19]  J. Scheller,et al.  ELPylated anti-human TNF therapeutic single-domain antibodies for prevention of lethal septic shock. , 2011, Plant biotechnology journal.

[20]  Natalya I. Topilina,et al.  Design and preparation of beta-sheet forming repetitive and block-copolymerized polypeptides. , 2007, Biomacromolecules.

[21]  Randolph V Lewis,et al.  Spider silk: ancient ideas for new biomaterials. , 2006, Chemical reviews.

[22]  Ashok Mulchandani,et al.  Temperature-triggered purification of antibodies. , 2005, Biotechnology and bioengineering.

[23]  Ashutosh Chilkoti,et al.  A thermally responsive biopolymer for intra-articular drug delivery. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[24]  Ashutosh Chilkoti,et al.  Stimulus-responsive macromolecules and nanoparticles for cancer drug delivery. , 2010, Nanomedicine.

[25]  J. Scheller,et al.  Functional expression of a biologically active fragment of soluble gp130 as an ELP-fusion protein in transgenic plants: purification via inverse transition cycling. , 2006, The Biochemical journal.

[26]  J. Scheller,et al.  Biochemical and functional characterization of anti-HIV antibody-ELP fusion proteins from transgenic plants. , 2008, Plant biotechnology journal.

[27]  A. Chilkoti,et al.  Purification of recombinant proteins from Escherichia coli at low expression levels by inverse transition cycling. , 2007, Analytical biochemistry.

[28]  H. Chu,et al.  A new cloning strategy for generating multiple repeats of a repetitive polypeptide based on non-template PCR , 2011, Biotechnology Letters.

[29]  C. Morrow,et al.  Production and Purification of a Recombinant Elastomeric Polypeptide, G‐(VPGVG)19‐VPGV, from Escherichia coli , 1992, Biotechnology progress.

[30]  Wilfred Chen,et al.  A temperature responsive biopolymer for mercury remediation. , 2003, Environmental science & technology.

[31]  Ashutosh Chilkoti,et al.  Elastin‐like polypeptides: Biomedical applications of tunable biopolymers , 2010, Biopolymers.

[32]  J. Rodríguez‐Cabello,et al.  Influence of the Molecular Weight on the Inverse Temperature Transition of a Model Genetically Engineered Elastin-like pH-Responsive Polymer , 2004 .

[33]  M. Yarmush,et al.  Modulation of single-chain antibody affinity with temperature-responsive elastin-like polypeptide linkers. , 2006, Biomacromolecules.

[34]  A. Chilkoti,et al.  Ultra‐High Expression of a Thermally Responsive Recombinant Fusion Protein in E. coli , 2006, Biotechnology progress.

[35]  Dan W. Urry,et al.  What Sustains Life?: Consilient Mechanisms for Protein-Based Machines and Materials , 2006 .

[36]  J. Scheller,et al.  Forcing single-chain variable fragment production in tobacco seeds by fusion to elastin-like polypeptides. , 2006, Plant biotechnology journal.

[37]  Wilfred Chen,et al.  Cadmium removal from contaminated soil by thermally responsive elastin (ELPEC20) biopolymers , 2007, Biotechnology and bioengineering.

[38]  M. Marquet,et al.  Genetic Engineering of Structural Protein Polymers , 1990, Biotechnology progress.

[39]  J. Rodríguez‐Cabello,et al.  Tailored recombinant elastin-like polymers for advanced biomedical and nano(bio)technological applications , 2006, Biotechnology Letters.

[40]  Margarida Casal,et al.  Proteolytic enzyme engineering: a tool for wool. , 2009, Biomacromolecules.

[41]  J. Fachet,et al.  Epitope Specificity of Monoclonal and Polyclonal Antibodies to Human Elastin , 1997, International Archives of Allergy and Immunology.

[42]  Ashutosh Chilkoti,et al.  Stimulus responsive elastin biopolymers: Applications in medicine and biotechnology. , 2006, Current opinion in chemical biology.

[43]  A. Weiss,et al.  Synthetic elastin hydrogels derived from massive elastic assemblies of self-organized human protein monomers. , 2004, Biomaterials.

[44]  A. Chilkoti,et al.  Protein Purification by Fusion with an Environmentally Responsive Elastin‐Like Polypeptide: Effect of Polypeptide Length on the Purification of Thioredoxin , 2001, Biotechnology progress.

[45]  Shuang-quan Zhang,et al.  Expression and purification of moricin CM4 and human β-defensins 4 in Escherichia coli using a new technology. , 2010, Microbiological research.

[46]  Felipe García Quiroz,et al.  Recursive directional ligation by plasmid reconstruction allows rapid and seamless cloning of oligomeric genes. , 2010, Biomacromolecules.

[47]  M. Dewhirst,et al.  Thermal cycling enhances the accumulation of a temperature-sensitive biopolymer in solid tumors. , 2007, Cancer research.

[48]  A. Fahmi,et al.  Nanopore formation by self-assembly of the model genetically engineered elastin-like polymer [(VPGVG)2(VPGEG)(VPGVG)2]15. , 2004, Journal of the American Chemical Society.

[49]  Wilfred Chen,et al.  Thermally triggered purification and immobilization of elastin–OPH fusions , 2003, Biotechnology and bioengineering.

[50]  Ashutosh Chilkoti,et al.  Synthesis and in vitro evaluation of enzymatically cross-linked elastin-like polypeptide gels for cartilaginous tissue repair. , 2005, Tissue engineering.

[51]  Ashutosh Chilkoti,et al.  Highly Parallel Method for Synthesis of DNA Repeats Enables Discovery of “Smart” Protein Polymers , 2010, Nature materials.

[52]  Teruyuki Nagamune,et al.  Recombinant extracellular matrix-like proteins with repetitive elastin or collagen-like functional motifs , 2005, Biotechnology Letters.

[53]  F Javier Arias,et al.  Emerging applications of multifunctional elastin-like recombinamers. , 2011, Nanomedicine.

[54]  H. Daniell,et al.  Expression of an environmentally friendly synthetic protein-based polymer gene in transgenic tobacco plants , 1996, Plant Cell Reports.

[55]  L. Setton,et al.  Fusion order controls expression level and activity of elastin‐like polypeptide fusion proteins , 2009, Protein science : a publication of the Protein Society.

[56]  Young Hwan Park,et al.  Native-sized recombinant spider silk protein produced in metabolically engineered Escherichia coli results in a strong fiber , 2010, Proceedings of the National Academy of Sciences.

[57]  V. Conticello,et al.  Rapid assembly of synthetic genes encoding protein polymers , 1999 .

[58]  Ashutosh Chilkoti,et al.  Self-assembling chimeric polypeptide-doxorubicin conjugate nanoparticles that abolish tumors after a single injection , 2009, Nature materials.

[59]  A. Taubert,et al.  Biomimetic calcium phosphate mineralization with multifunctional elastin-like recombinamers. , 2011, Biomacromolecules.

[60]  Ashutosh Chilkoti,et al.  Genetically encoded synthesis of protein-based polymers with precisely specified molecular weight and sequence by recursive directional ligation: examples from the elastin-like polypeptide system. , 2002, Biomacromolecules.

[61]  U. Conrad,et al.  High-Level Production and Long-Term Storage of Engineered Antibodies in Transgenic Tobacco Seeds , 1995, Bio/Technology.

[62]  C. García-Arévalo,et al.  Rapid micropatterning by temperature-triggered reversible gelation of a recombinant smart elastin-like tetrablock-copolymer , 2010 .

[63]  A. Chilkoti,et al.  Improved non-chromatographic purification of a recombinant protein by cationic elastin-like polypeptides. , 2007, Biomacromolecules.

[64]  E. Chaikof,et al.  Long-term biostability of self-assembling protein polymers in the absence of covalent crosslinking. , 2010, Biomaterials.

[65]  Javier Reguera,et al.  Biofunctional design of elastin-like polymers for advanced applications in nanobiotechnology , 2007, Journal of biomaterials science. Polymer edition.

[66]  M. Miao,et al.  Sequence and Structure Determinants for the Self-aggregation of Recombinant Polypeptides Modeled after Human Elastin* , 2003, Journal of Biological Chemistry.

[67]  A. Chilkoti,et al.  Morphing low-affinity ligands into high-avidity nanoparticles by thermally triggered self-assembly of a genetically encoded polymer. , 2010, ACS nano.

[68]  Rima Menassa,et al.  Optimization of elastin‐like polypeptide fusions for expression and purification of recombinant proteins in plants , 2009, Biotechnology and bioengineering.

[69]  H. Bayley,et al.  Temperature-responsive protein pores. , 2006, Journal of the American Chemical Society.

[70]  Raul Machado,et al.  Thermoresponsive self-assembled elastin-based nanoparticles for delivery of BMPs. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[71]  A. Hatefi,et al.  Advances in recombinant polymers for delivery of bioactive agents. , 2010, Advanced drug delivery reviews.

[72]  Jay D Keasling,et al.  Improved assembly of multimeric genes for the biosynthetic production of protein polymers. , 2002, Biomacromolecules.

[73]  Jie Xu,et al.  Product purification by reversible phase transition following Escherichia coli expression of genes encoding up to 251 repeats of the elastomeric pentapeptide GVGVP. , 1996, Protein expression and purification.

[74]  Byung-Gee Kim,et al.  The effects of supplementing specific amino acids on the expression of elastin-like polypeptides (ELPs). , 2010, Protein expression and purification.

[75]  E. Chaikof,et al.  Expression of a recombinant elastin‐like protein in pichia pastoris , 2009, Biotechnology progress.

[76]  J. Rodríguez‐Cabello,et al.  Influence of the amino-acid sequence on the inverse temperature transition of elastin-like polymers. , 2009, Biophysical journal.

[77]  S. Jana,et al.  RETRACTED ARTICLE: Strategies for efficient production of heterologous proteins in Escherichia coli , 2005, Applied Microbiology and Biotechnology.

[78]  C. Filipe,et al.  Simultaneous phase transition of ELP tagged molecules and free ELP: an efficient and reversible capture system. , 2006, Biomacromolecules.

[79]  G. Eggink,et al.  Secreted production of an elastin-like polypeptide by Pichia pastoris , 2009, Applied Microbiology and Biotechnology.

[80]  Wilfred Chen,et al.  Tunable Biopolymers for Heavy Metal Removal , 2001 .

[81]  Liang Feng,et al.  Simple bioseparations using self-cleaving elastin-like polypeptide tags , 2005, Nature Methods.

[82]  J. Rodríguez‐Cabello,et al.  Genetic Engineering of Protein-Based Polymers: The Example of Elastinlike Polymers , 2005 .

[83]  Ashutosh Chilkoti,et al.  Fabrication of a reversible protein array directly from cell lysate using a stimuli-responsive polypeptide. , 2003, Analytical chemistry.

[84]  Annelise E. Barron,et al.  A New Cloning Method for the Preparation of Long Repetitive Polypeptides without a Sequence Requirement , 2002 .

[85]  Wenxin Wang,et al.  Tunable elastin-like polypeptide hollow sphere as a high payload and controlled delivery gene depot. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[86]  V. Conticello,et al.  Self-assembly of block copolymers derived from elastin-mimetic polypeptide sequences. , 2002, Advanced drug delivery reviews.

[87]  Lixin Mi Molecular cloning of protein-based polymers. , 2006, Biomacromolecules.

[88]  Wilfred Chen,et al.  Affinity purification of plasmid DNA by temperature-triggered precipitation , 2007, Nature Protocols.

[89]  Eddie Wang,et al.  Elastin-like polypeptide based hydroxyapatite bionanocomposites. , 2011, Biomacromolecules.

[90]  Brigida Bochicchio,et al.  Investigating by CD the molecular mechanism of elasticity of elastomeric proteins. , 2008, Chirality.

[91]  T. LaBean,et al.  Effect of protein fusion on the transition temperature of an environmentally responsive elastin-like polypeptide: a role for surface hydrophobicity? , 2004, Protein engineering, design & selection : PEDS.

[92]  A. Weiss,et al.  Cellular interactions with elastin. , 2005, Pathologie-biologie.