Array-Based High-Throughput Analysis of Silk-Elastinlike Protein Polymer Degradation and C-Peptide Release by Proteases

  • Anal Chem. 2016 May 17;88(10):5398-405. doi: 10.1021/acs.analchem.6b00739.
Hye-Yoon Jeon  1 ,  Se-Hui Jung  1 ,  Young Mee Jung  2 ,  Young-Myeong Kim  1 ,  Hamidreza Ghandehari  3  4 ,  Kwon-Soo Ha  1
Affiliations
  • 1. Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine , Chuncheon, Kangwon-Do 200-701, Korea.
  • 2. Department of Chemistry, Kangwon National University , Chuncheon, Kangwon-Do 200-701, Korea.
  • 3. Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology , Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Korea.
  • 4. Departments of Phamaceutics and Pharmaceutical Chemistry, and Bioengineering, Center for Nanomedicine, Nano Institute of Utah, University of Utah , Salt Lake City, Utah 84112, United States.
PMID: 27109435 DOI: 10.1021/acs.analchem.6b00739
Abstract

The objective of this study was to utilize an on-chip degradation assay to evaluate polymer depots and the predicted drug release from the depots. We conjugated four silk-elastinlike protein (SELP) Polymers including SELP-815K, SELP-815K-RS1, SELP-815K-RS2, and SELP-815K-RS5 with a Cy5-NHS ester and fabricated SELP arrays by immobilizing the conjugated Polymers onto well-type amine arrays. SELP polymer degradation rates were investigated by calculating the half-maximal effective concentration (EC50). Eight cleavage Enzymes were applied, all of which exhibited distinctive EC50 values for SELP-815K and its three analogues. We successfully utilized this assay to study the in vitro release of the Cy5-conjugated C-peptide from SELP-815K hydrogel arrays. Additionally, cumulative C-peptide release from the SELP-815K depots was also demonstrated using repetitive Elastase treatments. Therefore, this array-based on-chip degradation assay could potentially be used for evaluating depot degradation and controlled drug release from polymer depots at the molecular level.

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