Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.
Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.
Anal Chim Acta. 2022 Apr 29;1204:339697. doi: 10.1016/j.aca.2022.339697. Epub 2022 Mar 9.
In this study, a strategy of improving imprinting performance was developed using an enhanced cooperation effect of functional monomers based on deep eutectic solvents (DESs) monomer for the specific enrichment of benzoylation modified peptides. Zinc acrylate and DESs monomers were used as binary functional monomers, and ethylene glycol dimethacrylate was used as the cross-linking agent with SGRGKbz as template to prepare an imprinted monolith. It was observed that the use of DESs monomer significantly improveed the affinity of benzoylation imprinted monolith and increased the adsorption capacity. Under optimal conditions, the recovery and imprinting factor (IF) of the imprinted monolith for SGRGKbz can reach 93.0% and 10.58, respectively. The average recovery of SGRGKbz extracted from the spiked histone digestion solution can reach 88.4% (n = 5, RSD = 3.4%). After treatment with the benzoylation imprinted monolith, 12 benzoylation modified peptides, 13 benzoylation modified sites and 12 benzoylation proteins could be identified in the digestion of mouse liver protein, while only one of each benzoylation modified peptide, benzoylation modified site and benzoylation protein could be identified in the untreated digestion of mouse liver protein. The results indicated that the prepared imprinted monolith using DESs-based functional monomer was an effective method to increase the affinity of the resulting MIP.
在这项研究中，开发了一种基于深共熔溶剂（DESs）单体的功能单体增强协同作用的策略，用于特异性富集苯甲酰化修饰肽。使用丙烯酸钠和 DESs 单体作为二元功能单体，乙二醇二甲基丙烯酸酯作为交联剂，以 SGRGKbz 为模板制备印迹整体柱。结果表明，DESs 单体的使用显著提高了苯甲酰化印迹整体柱的亲和力，并增加了吸附容量。在最佳条件下，印迹整体柱对 SGRGKbz 的回收率和印迹因子（IF）分别可达 93.0%和 10.58。从掺杂组蛋白消化液中提取的 SGRGKbz 的平均回收率可达 88.4%（n=5，RSD=3.4%）。经苯甲酰化印迹整体柱处理后，可从小鼠肝蛋白消化物中鉴定出 12 种苯甲酰化修饰肽、13 个苯甲酰化修饰位点和 12 种苯甲酰化蛋白，而未经处理的小鼠肝蛋白消化物中仅能鉴定出每种苯甲酰化修饰肽、苯甲酰化修饰位点和苯甲酰化蛋白各一种。结果表明，使用基于 DESs 的功能单体制备的印迹整体柱是一种有效提高所得 MIP 亲和力的方法。
