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

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• 作者：Hye-Yoon Jeon ; Se-Hui Jung ; Young Mee Jung ; Young-Myeong Kim ; Hamidreza Ghandehari ; Kwon-Soo Ha
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：May 17, 2016
• 年：2016
• 卷：88
• 期：10
• 页码：5398-5405
• 全文大小：522K
• 年卷期：0
• ISSN：1520-6882

文摘

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.