The overall objective of this competing R01 renewal is the continued development of the Elastin Fusion Protein technology as a genetically encodable, biomimetic analog of -""""""""smart"""""""" protein-polymer conjugates. The overall objective is motivated by the rationale that simple and convenient methods to modulate the physico-chemical properties of proteins -most notably their solubility in solution and binding to a surface- in response to an external trigger have immediate application in many areas of Bioengineering and Biotechnology such as protein purification, affinity capture, protein chips, lab-on-a-chip bioanalysis and in the development of environmentally responsive biomaterials and nanocarriers for delivery of protein therapeutics. The objective of the proposed research will be accomplished by the gene-level fusion of different recombinant proteins to a stimulus-responsive elastin-like polypeptide (ELP) tag to create ELP fusion proteins. The overall hypothesis of this research -validated for a set of proteins in the previous funding period- is that environmentally sensitive and reversible solubility and binding to a surface can be imparted to any soluble protein of interest by N-or C-terminal fusion to an ELP sequence. The proposed research has two research foci: the first, """"""""fundamental focus"""""""", seeks to discover the biophysical basis of the effect that fusion of proteins has on the inverse transition behavior of an ELP. The molecular level understanding of the inverse transition behavior of ELP fusion proteins will directly feed into the second, """"""""technological focus"""""""" that will develop an ensemble of applications that exploit the inverse transition behavior of ELP fusion proteins. The second focus consists of three related technological applications: (1) optimization of the expression and purification of ELP fusion proteins in a bacterial expression system; (2) affinity capture of biomolecules by an ELP fusion protein followed by their separation by inverse transition cycling; and (3) """"""""direct, on chip"""""""", reversible capture of ELP fusion proteins from solution on surfaces functionalized with ELPs. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM061232-08
Application #
7392840
Study Section
Special Emphasis Panel (ZRG1-BMBI (01))
Program Officer
Smith, Ward
Project Start
2000-04-01
Project End
2009-07-31
Budget Start
2008-04-01
Budget End
2009-07-31
Support Year
8
Fiscal Year
2008
Total Cost
$242,718
Indirect Cost
Name
Duke University
Department
Biomedical Engineering
Type
Schools of Engineering
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Roberts, Stefan; Harmon, Tyler S; Schaal, Jeffrey L et al. (2018) Author Correction: Injectable tissue integrating networks from recombinant polypeptides with tunable order. Nat Mater 17:1164
Costa, Simone A; Simon, Joseph R; Amiram, Miriam et al. (2018) Photo-Crosslinkable Unnatural Amino Acids Enable Facile Synthesis of Thermoresponsive Nano- to Microgels of Intrinsically Disordered Polypeptides. Adv Mater 30:
Li, Linying; Li, Nan K; Tu, Qing et al. (2018) Functional Modification of Silica through Enhanced Adsorption of Elastin-Like Polypeptide Block Copolymers. Biomacromolecules 19:298-306
Dzuricky, Michael; Roberts, Stefan; Chilkoti, Ashutosh (2018) Convergence of Artificial Protein Polymers and Intrinsically Disordered Proteins. Biochemistry 57:2405-2414
Roberts, Stefan; Harmon, Tyler S; Schaal, Jeffrey L et al. (2018) Injectable tissue integrating networks from recombinant polypeptides with tunable order. Nat Mater 17:1154-1163
Gilroy, Caslin A; Roberts, Stefan; Chilkoti, Ashutosh (2018) Fusion of fibroblast growth factor 21 to a thermally responsive biopolymer forms an injectable depot with sustained anti-diabetic action. J Control Release 277:154-164
Gonzalez, Mark A; Simon, Joseph R; Ghoorchian, Ali et al. (2017) Strong, Tough, Stretchable, and Self-Adhesive Hydrogels from Intrinsically Unstructured Proteins. Adv Mater 29:
Luginbuhl, Kelli M; Mozhdehi, Davoud; Dzuricky, Michael et al. (2017) Recombinant Synthesis of Hybrid Lipid-Peptide Polymer Fusions that Self-Assemble and Encapsulate Hydrophobic Drugs. Angew Chem Int Ed Engl 56:13979-13984
Weitzhandler, Isaac; Dzuricky, Michael; Hoffmann, Ingo et al. (2017) Micellar Self-Assembly of Recombinant Resilin-/Elastin-Like Block Copolypeptides. Biomacromolecules 18:2419-2426
Simon, Joseph R; Carroll, Nick J; Rubinstein, Michael et al. (2017) Programming molecular self-assembly of intrinsically disordered proteins containing sequences of low complexity. Nat Chem 9:509-515

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