Zhang Ka, Zhou Tongchang, Dicko Cedric, Ye Lei, Bülow Leif
Division of Pure and Applied Biochemistry, Department of Chemistry, Lund University, Box 124, 22100 Lund, Sweden.
Polymers (Basel). 2024 Sep 27;16(19):2734. doi: 10.3390/polym16192734.
Development in hemoglobin-based oxygen carriers (HBOCs) that may be used as alternatives to donated blood requires an extensive supply of highly pure hemoglobin (Hb) preparations. Therefore, it is essential to fabricate inexpensive, stable and highly selective absorbents for Hb purification. Molecular imprinting is an attractive technology for preparing such materials for targeted molecular recognition and rapid separations. In this case study, we developed human fetal hemoglobin (HbF)-imprinted polymer beads through the fusion of surface imprinting and Pickering emulsion polymerization. HbF was firstly covalently coupled to silica nanoparticles through its surface-exposed amino groups. The particle-supported HbF molecules were subsequently employed as templates for the synthesis of molecularly imprinted polymers (MIPs) with high selectivity for Hb. After removing the silica support and HbF, the resulting MIPs underwent equilibrium and kinetic binding experiments with both adult Hb (HbA) and HbF. These surface-imprinted MIPs exhibited excellent selectivity for both HbA and HbF, facilitating the one-step isolation of recombinant Hb from crude biological samples. The saturation capacities of HbA and HbF were found to be 15.4 and 17.1 mg/g polymer, respectively. The present study opens new possibilities for designed resins for tailored protein purification, separation and analysis.
开发可作为捐献血液替代品的基于血红蛋白的氧载体(HBOCs)需要大量高纯度血红蛋白(Hb)制剂。因此,制造用于Hb纯化的廉价、稳定且高选择性的吸附剂至关重要。分子印迹是一种用于制备此类具有靶向分子识别和快速分离功能材料的有吸引力的技术。在本案例研究中,我们通过表面印迹和Pickering乳液聚合相结合的方法,制备了人胎儿血红蛋白(HbF)印迹聚合物微球。首先,HbF通过其表面暴露的氨基与二氧化硅纳米颗粒共价偶联。随后,以负载在颗粒上的HbF分子为模板,合成了对Hb具有高选择性的分子印迹聚合物(MIPs)。去除二氧化硅载体和HbF后,所得MIPs与成人血红蛋白(HbA)和HbF进行了平衡和动力学结合实验。这些表面印迹的MIPs对HbA和HbF均表现出优异的选择性,有助于从粗生物样品中一步分离重组Hb。发现HbA和HbF的饱和容量分别为15.4和17.1 mg/g聚合物。本研究为设计用于定制蛋白质纯化、分离和分析的树脂开辟了新的可能性。