Liu Peiji, Lin Hong, Wang Jingxue
College of Food Science and Engineering, Ocean University of China, Huangdao Campus, Qingdao 266400, China.
Gels. 2025 May 28;11(6):402. doi: 10.3390/gels11060402.
To address the limitations of traditional endotoxin adsorbents, which exhibit poor endotoxin removal efficiency and low sample recovery when processing high-concentration samples, a novel cryogel, CG(HEMA-co-AM), based on acrylamide (AM) and hydroxyethyl methacrylate (HEMA) as the second monomer, was successfully designed and synthesized. After optimizing the epoxidation and polymyxin B (PMB) conjugation processes, leading to the successful preparation of the functionalized cryogel CG(HEMA-co-AM)@ECH@PMB, flow-through experiments showed that in Tris-HCl buffer at a flow rate of 6 mL/min, the endotoxin removal efficiency reached 99.82%, with a maximum adsorption capacity of 1408.38 EU/mg. In a complex protein system containing BSA, HSA, Hb, LYS, and OVA (each at 10 mg/mL), the maximum endotoxin removal efficiency was 99.62%. Further investigations revealed that pH and ionic strength critically influenced the regulation of hydrophobic and electrostatic interactions, thus significantly affecting the endotoxin adsorption efficiency. Additionally, weakly hydrophobic and basic lysozyme exhibited significantly higher recovery rates compared to strongly hydrophobic and acidic proteins such as BSA and OVA. This functionalized cryogel integrates a high adsorption capacity with excellent specificity and features a macroporous structure suitable for dynamic chromatographic separation. It offers a novel, reusable adsorbent material for endotoxin removal in protein preparation, biopharmaceutical production, and clinical blood purification applications.
为解决传统内毒素吸附剂的局限性，即在处理高浓度样品时内毒素去除效率低且样品回收率低的问题，一种基于丙烯酰胺（AM）和甲基丙烯酸羟乙酯（HEMA）作为第二单体的新型冷冻凝胶CG（HEMA-co-AM）被成功设计并合成。在优化环氧化和多粘菌素B（PMB）偶联过程后，成功制备了功能化冷冻凝胶CG（HEMA-co-AM）@ECH@PMB，流通实验表明，在Tris-HCl缓冲液中，流速为6 mL/min时，内毒素去除效率达到99.82%，最大吸附容量为1408.38 EU/mg。在含有牛血清白蛋白（BSA）、人血清白蛋白（HSA）、血红蛋白（Hb）、赖氨酸（LYS）和卵清蛋白（OVA）（各10 mg/mL）的复杂蛋白质体系中，最大内毒素去除效率为99.62%。进一步研究表明，pH值和离子强度对疏水和静电相互作用的调节有至关重要的影响，从而显著影响内毒素吸附效率。此外，与BSA和OVA等强疏水和酸性蛋白质相比，弱疏水和碱性的溶菌酶表现出明显更高的回收率。这种功能化冷冻凝胶兼具高吸附容量和优异的特异性，具有适合动态色谱分离的大孔结构。它为蛋白质制备、生物制药生产和临床血液净化应用中的内毒素去除提供了一种新型的、可重复使用的吸附材料。
