Array-Based High-Throughput Analysis of Silk-Elastinlike Protein Polymer Degradation and C-Peptide Release by Proteases.
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.
Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine , Chuncheon, Kangwon-Do 200-701, Korea.
Hye-Yoon
HY
Jeon
Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine , Chuncheon, Kangwon-Do 200-701, Korea.
Se-Hui
SH
Jung
Department of Chemistry, Kangwon National University , Chuncheon, Kangwon-Do 200-701, Korea.
Young Mee
YM
Jung
Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine , Chuncheon, Kangwon-Do 200-701, Korea.
Young-Myeong
YM
Kim
Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology , Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Korea.|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.
Hamidreza
H
Ghandehari
Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine , Chuncheon, Kangwon-Do 200-701, Korea.
Kwon-Soo
KS
Ha
D000595:Amino Acid Sequence; D002096:C-Peptide / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D002232:Carbocyanines / Q000737:chemistry; D057166:High-Throughput Screening Assays; D020100:Hydrogels / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D010447:Peptide Hydrolases / Q000378:metabolism*; D047011:Silk / Q000737:chemistry / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared
D002096:C-Peptide; D002232:Carbocyanines; D020100:Hydrogels; D047011:Silk; C085321:cyanine dye 5; D010447:Peptide Hydrolases