Muntiza Nurimangul, Zhang Wenbin, Li Yuzeng, Ji Wenquan, Zhao Jin, Zhang Hongfeng, Li Qinran, Deng Qiliang, Sun Donglan, Liu Tianjun
College of Chemical Engineering and Materials Science, Tianjin University of Science and Technology, Tianjin, 300457, People's Republic of China; College of Science, Tianjin University of Science and Technology, Tianjin, 300457, People's Republic of China.
College of Chemical Engineering and Materials Science, Tianjin University of Science and Technology, Tianjin, 300457, People's Republic of China.
Talanta. 2025 May 1;286:127567. doi: 10.1016/j.talanta.2025.127567. Epub 2025 Jan 10.
A novel strategy for cytochrome c selective recognition assisted with cucurbit[6]uril by host-guest interaction via N-terminal epitope imprinting and reversible addition-fragmentation chain transfer (RAFT) polymerization was developed. N-terminal nonapeptide of cytochrome c (GI-9) was used as the epitope template to achieve highly selective recognition of cytochrome c. As a common supramolecule in recent years, cucurbit[6]uril can encapsulate the butyrammonium group of lysine residue to capture the peptide and improve the corresponding spatial orientation by the host-guest interaction for GI-9 or cytochrome c recognition. After cucurbit[6]uril modification and epitope immobilization, the imprinted polymer was synthesized by RAFT polymerization with 2-dodecylsulfanylcarbothioylsulfanyl-2-methylpropanoic acid as chain transfer agent. After template removal, the obtained imprinted particles showed good binding ability to GI-9 (20.28 mg g, IF = 4.11) and cytochrome c (36.12 mg g, IF = 3.91). With the successive addition of cucurbit[6]uril and RAFT agent, the step-by-step improvement of the IF for cytochrome c recognition further illustrated the effects of supramolecular host-guest interaction and regulation of imprinted polymer chain. The imprinted polymers showed obvious advantages for cytochrome c recognition compared to competitive proteins and had good reusability with the repeated reproduction rate 80.8 % after five cycles of adsorption and desorption. Furthermore, the selective recognition for cytochrome c in adult bovine serum proved its potentiality to be applied in practical samples. All these results demonstrated that the combination of epitope imprinting, cucurbit[6]uril host-guest interaction and RAFT strategy presented an efficient new feasible control method for protein recognition with good selectivity, stability and reusability.
通过主客体相互作用，借助葫芦[6]脲，采用N端表位印迹和可逆加成-断裂链转移（RAFT）聚合反应，开发了一种用于细胞色素c选择性识别的新策略。细胞色素c的N端九肽（GI-9）用作表位模板，以实现对细胞色素c的高度选择性识别。作为近年来常见的超分子，葫芦[6]脲可以包裹赖氨酸残基的丁铵基团，以捕获肽，并通过主客体相互作用改善相应的空间取向，用于GI-9或细胞色素c的识别。经过葫芦[6]脲修饰和表位固定后，以2-十二烷基硫烷基硫代羰基硫烷基-2-甲基丙酸为链转移剂，通过RAFT聚合反应合成了印迹聚合物。去除模板后，所得印迹颗粒对GI-9（20.28 mg g，IF = 4.11）和细胞色素c（36.12 mg g，IF = 3.91）表现出良好的结合能力。随着葫芦[6]脲和RAFT试剂的相继加入，细胞色素c识别的IF逐步提高，进一步说明了超分子主客体相互作用和印迹聚合物链调控的效果。与竞争性蛋白质相比，印迹聚合物在细胞色素c识别方面具有明显优势，并且在经过五次吸附和解吸循环后，具有良好的可重复使用性，重复再生率为80.8%。此外，在成年牛血清中对细胞色素c的选择性识别证明了其应用于实际样品的潜力。所有这些结果表明，表位印迹、葫芦[6]脲主客体相互作用和RAFT策略的结合为蛋白质识别提供了一种高效、新的可行控制方法，具有良好的选择性、稳定性和可重复使用性。
