In solid-phase peptide synthesis (SPPS), a high concentration of functional groups on the solid support and an adequate swelling volume enables its use for high-throughput screening of receptor agonists and therapeutic peptides. However, a high resin loading often leads to insufficient purity of the synthesized peptides and false positive binding with the target due to improper interactions between neighboring peptides. Therefore, this study focused on low-loaded polyethylene glycol (PEG) resins to achieve high-specificity screening using a core–shell-type PEG hydrogel resin. The peptide purity and target specificity were determined by assessing (1) the resin swelling properties in various solvents, (2) purity of a complicated Jung-Redemann (JR) decapeptide, and (3) the cell-adhesive behavior of GRGDS-pentapeptide-bound resins. The results were compared with those obtained using polyacrylamide resin (PAM) and conventionally used TentaGel S NH₂ resin (TG^®). The highest JR decapeptide purity was achieved using the PEG-based resin with a higher degree of cross-linking (PEGHN). Furthermore, the resin was preferably qualified as an extracellular microenvironment to accommodate true specific binding with fibroblast cells. Thus, SPPS and cell binding assays using the developed PEG-based resin provide a novel stringent strategy with potential application for true positive screening in biological assays.

在固相肽合成（SPPS）中，固体支持物上的高浓度官能团和足够的溶胀体积使其能够用于受体激动剂和治疗性肽的高通量筛选。然而，高树脂负载量通常会导致合成肽的纯度不足，并且由于相邻肽之间的不正确相互作用而导致与靶标的结合出现假阳性。因此，本研究重点关注低负载量的聚乙二醇（PEG）树脂，以利用核壳型PEG水凝胶树脂实现高特异性筛选。通过评估（1）树脂在各种溶剂中的溶胀特性，（2）复杂的Jung-Redemann（JR）十肽的纯度，以及（3）GRGDS-五肽-的细胞粘附行为来确定肽纯度和靶标特异性。结合树脂。将结果与使用聚丙烯酰胺树脂(PAM)和传统使用的TentaGel S NH ₂树脂(TG ^® )获得的结果进行比较。使用具有较高交联度的 PEG 树脂 (PEGHN) 实现了最高的 JR 十肽纯度。此外，树脂优选适合作为细胞外微环境，以适应与成纤维细胞的真正特异性结合。因此，使用开发的基于 PEG 的树脂进行 SPPS 和细胞结合测定提供了一种新颖的严格策略，具有在生物测定中真正阳性筛选的潜在应用。