June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Bridging ocular therapeutics and contact lenses via thermo-responsive protein polymers
Author Affiliations & Notes
  • Wan Wang
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
  • Pu Shi
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
  • Suhaas Aluri
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
  • Pang-Yu Hsueh
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
  • Maria Edman
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
  • Denise Ryan
    US Army Warfighter Refractive Surg Research Ctr, Fort Belvoir, VA
  • Robert McKown
    Department of Integrated Science and Technology, James Madison University, Harrisonburg, VA
  • Sarah Hamm-Alvarez
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
    Keck School of Medicine, University of Southern California, Los Angeles, CA
  • Gordon Laurie
    University of Virginia, Charlottesville, VA
  • John MacKay
    Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA
    Department of Biomedical Engineering, University of Southern California, Los Angeles, CA
  • Footnotes
    Commercial Relationships Wan Wang, University of Southern California (P); Pu Shi, None; Suhaas Aluri, None; Pang-Yu Hsueh, University of Southern California (P); Maria Edman, University of Southern California (P); Denise Ryan, None; Robert McKown, EyeRx Research, Inc. (I); Sarah Hamm-Alvarez, None; Gordon Laurie, UVa Patent Foundation (F); John MacKay, University of Southern California (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2668. doi:
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      Wan Wang, Pu Shi, Suhaas Aluri, Pang-Yu Hsueh, Maria Edman, Denise Ryan, Robert McKown, Sarah Hamm-Alvarez, Gordon Laurie, John MacKay, ; Bridging ocular therapeutics and contact lenses via thermo-responsive protein polymers. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2668.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: To influence the bioavailability of the prosecretory mitogen lacritin, we fused it with elastin-derived protein polymers, elastin-like polypeptides (ELPs) that have a range of assembly properties. ELPs reversibly phase separated onto ProclearTM contact lens and may provide an alternative route for delivery of protein therapeutics to the ocular surface system.

Methods: Lac-ELP genes were expressed in E. coli and purified utilizing Inverse Transition Cycling (ITC) followed by size exclusion chromatography. Phase behavior and nanoparticle formation of fusion proteins were characterized by optical density, Dynamic Light Scattering (DLS), TEM and Cryo-TEM. In vitro activities were tested using rabbit lacrimal gland acinar cells (LGACs) and SV40-immortalized human corneal epithelial cells (HCE-T). In vitro ProclearTM contact lens retention was characterized by fluorescence detection.

Results: A novel fusion protein library based on recombinant human lacritin and elastin-like-polypeptides (ELPs) was designed as a potential therapeutic for the ocular surface. The Lacritin-ELP fusion proteins imparted thermo-sensitive assembly of micron-sized coacervates or 130-140nm diameter micelles depending on the sequence of their ELP tag. Exogenous Lac-ELPs promoted β-hexosaminidase secretion from rabbit lacrimal gland acinar cells (LGACs); evoked Ca2+ wave propagation across Human Corneal Epithelial Cells (HCE-T) and were taken up by both LGACs and HCE-T cells. ELPs reversibly attached onto ProclearTM contact lens and the total retention time was temperature and Tt dependent.

Conclusions: Exploration of thermo-responsive prosecretory mitogen lacritin-ELPs may provide an alternative approach to adjust their ocular retention and interaction with target cellular receptor. Moreover, biocompatible elastin-like polypeptides (ELPs) appear to provide a reversible, temperature dependent bridge between potential ocular therapeutics and other polymers used on the ocular surface.

Keywords: 486 cornea: tears/tear film/dry eye • 765 wound healing • 477 contact lens  
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