Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto, Japan.
Department of Chemistry and Biomolecular Science, Faculty of Engineering, Gifu University, Gifu, Japan.
Nat Protoc. 2016 Sep;11(9):1744-56. doi: 10.1038/nprot.2016.099. Epub 2016 Aug 25.
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.
水凝胶因子是小分子，能够自组装形成超分子纳米纤维网络，表现出独特的动态特性。开发能够响应各种生物分子降解的超分子水凝胶，可能会被应用于药物输送和诊断等领域。在此，我们提供了一种制备氧化还原响应性超分子水凝胶因子的合成方法，用于制备能够响应氧化或还原条件而降解的水凝胶。该合成过程大约需要 2-4 天，并且可以并行进行，以制备多个水凝胶因子候选物。本文描述的固相肽合成方案可用于制备先前描述的水凝胶因子，或构建一个聚焦的分子文库，以有效地发现和优化新的水凝胶因子。此外，我们还描述了氧化还原响应性超分子水凝胶-酶杂合体的制备方法，将水凝胶因子和酶的水溶液混合即可完成制备，整个过程需要 2 小时。所得的超分子水凝胶-酶杂合体可以响应各种生物分子进行凝胶降解，并且可以通过将水凝胶因子的化学反应与酶反应连接起来进行合理设计。响应生物分子作为触发因素的凝胶降解在数小时内发生。我们还描述了在平玻璃片上排列的水凝胶-酶杂合体的制备方法，通过凝胶降解可以实现对葡萄糖、尿酸、乳酸等生物分子的高通量分析，通过肉眼即可检测到。该方案大约需要 6 小时来制备水凝胶-酶杂合体阵列并完成生物分子分析。
