title: Preparation of supramolecular hydrogel-enzyme hybrids exhibiting biomolecule-responsive gel degradation.
abstract: Hydrogelators are small, self-assembling molecules that form supramolecular nanofiber networks that exhibit unique dynamic properties. Development of supramolecular hydrogels that degrade in response to various biomolecules could potentially be used for applications in areas such as drug delivery and diagnostics. Here we provide a synthetic procedure for preparing redox-responsive supramolecular hydrogelators that are used to create hydrogels that degrade in response to oxidizing or reducing conditions. The synthesis takes ∼2-4 d, and it can potentially be carried out in parallel to prepare multiple hydrogelator candidates. This described solid-phase peptide synthesis protocol can be used to produce previously described hydrogelators or to construct a focused molecular library to efficiently discover and optimize new hydrogelators. In addition, we describe the preparation of redox-responsive supramolecular hydrogel-enzyme hybrids that are created by mixing aqueous solutions of hydrogelators and enzymes, which requires 2 h for completion. The resultant supramolecular hydrogel-enzyme hybrids exhibit gel degradation in response to various biomolecules, and can be rationally designed by connecting the chemical reactions of the hydrogelators with enzymatic reactions. Gel degradation in response to biomolecules as triggers occurs within a few hours. We also describe the preparation of hydrogel-enzyme hybrids arrayed on flat glass slides, enabling high-throughput analysis of biomolecules such as glucose, uric acid, lactate and so on by gel degradation, which is detectable by the naked eye. The protocol requires ∼6 h to prepare the hydrogel-enzyme hybrid array and to complete the biomolecule assay.
authors: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.
authors: Hajime
authors: H
authors: Shigemitsu
authors: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.
authors: Takahiro
authors: T
authors: Fujisaku
authors: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.
authors: Shoji
authors: S
authors: Onogi
authors: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.
authors: Tatsuyuki
authors: T
authors: Yoshii
authors: Department of Chemistry and Biomolecular Science, Faculty of Engineering, Gifu University, Gifu, Japan.|Department of Biomolecular Science, Graduate School of Engineering, Gifu University, Gifu, Japan.|United Graduate School of Drug Discovery and Medical Information Sciences, Gifu University, Gifu, Japan.
authors: Masato
authors: 0000-0003-4097-8292
authors: M
authors: Ikeda
authors: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.|Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo, Japan.
authors: Itaru
authors: I
authors: Hamachi
mesh_terms: D060326:Chemistry Techniques, Synthetic / Q000379:methods*; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism; D020100:Hydrogels / Q000138:chemical synthesis* / Q000737:chemistry*; D007700:Kinetics; D010084:Oxidation-Reduction; D014867:Water / Q000737:chemistry
chemical_list: D004798:Enzymes; D020100:Hydrogels; D014867:Water