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基于自聚合二氧化硅纳米粒子的分子印迹聚合物用于蛋白质的选择性识别。

Self-polymerization silica nanoparticles based molecularly imprinted polymers for selective recognition of protein.

机构信息

Department of Pharmaceutical Analysis, School of Pharmacy, Key Laboratory of Protection, Development and Utilization of Medicinal Resources in Liupanshan Area, Ministry of Education, Ningxia Medical University, Yinchuan 750004, China.

Center for Hybrid Nanostructures, Universität Hamburg, Hamburg, Germany.

出版信息

J Chromatogr A. 2024 Sep 13;1732:465260. doi: 10.1016/j.chroma.2024.465260. Epub 2024 Aug 12.

Abstract

Molecularly imprinted polymers (MIPs) are promising for precise protein separation and purification. However, challenges persist due to their large size, variable configuration, and instability during preparation. Here, a simple silicon self-assembly program was designed to synthesize MIPs without any organic reagents and acid-base catalysis, avoiding the structural damage of protein under severe conditions. In this method, employing hemoglobin (Hb) as a model protein, with tween-20 in emulsification, and tetraethyl orthosilicate (TEOS) as the cross-linking agent, along with co-functional monomers 3-aminopropyltriethoxysilane (APTES) and benzyl(triethoxy)silane (BnTES), enhanced binding efficacy was achieved. Successful imprinting was evidenced through surface morphology observation and physical/chemical property evaluations of the synthesized MIPs. A series of adsorption experiments were performed to investigate the recognition performance of Hb-MIPs. The Hb-MIPs not only exhibited large adsorption capacity (400 μg/mg) and good imprinting factor (6.09) toward template protein, but also showed satisfactory selectivity for reference proteins. Five cycles of adsorption proved that the Hb-MIPs had good reusability. In addition, the successful isolation of HB from bovine blood indicated that Hb-MIPs were an excellent separation and purification material. The mild preparation conditions and good adsorption capacity demonstrated the potential value of this method in separation and purification research.

摘要

分子印迹聚合物(MIPs)在精确的蛋白质分离和纯化方面具有很大的应用前景。然而,由于其尺寸较大、结构多变且在制备过程中不稳定,仍然存在挑战。在此,设计了一种简单的硅自组装程序,用于合成 MIPs,无需任何有机试剂和酸碱催化,避免了蛋白质在苛刻条件下的结构损伤。在该方法中,以血红蛋白(Hb)为模型蛋白,使用吐温-20 乳化,正硅酸乙酯(TEOS)作为交联剂,同时使用共功能单体 3-氨丙基三乙氧基硅烷(APTES)和苄基(三乙氧基)硅烷(BnTES),提高了结合效率。通过对合成的 MIPs 的表面形貌观察和物理/化学性质评估,证明了成功的印迹。进行了一系列吸附实验来研究 Hb-MIPs 的识别性能。Hb-MIPs 不仅对模板蛋白表现出较大的吸附容量(400μg/mg)和良好的印迹因子(6.09),而且对参考蛋白也表现出良好的选择性。五次吸附循环证明了 Hb-MIPs 具有良好的可重复使用性。此外,从牛血中成功分离出 HB 表明 Hb-MIPs 是一种优秀的分离和纯化材料。温和的制备条件和良好的吸附容量表明该方法在分离和纯化研究中具有潜在的应用价值。

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