Abstract Purification and analyzing of proteins is an essential means for understanding their function and diseases associated with their lack or defect. In this research, a new lanthanide-chelate based molecularly imprinted polymer (MIP) was synthesized for selective separation of Hemoglobin (Hb) from human serum in the presence of various interference molecules. The Hb-imprinted polymer was prepared by using complex functional monomer N-methacryloylamido antipyrine (MAAP)-Ce(III) and 2-Hydroxyethyl methacrylate (HEMA) in accordance of cryopolymerization techniques. The nonimprinted cryogel (NIP) was also prepared at same polymerization conditions in the absence of template Hb molecule. The effects of pH, initial Hb concentration, flow rate, temperature and ionic strength on the binding capacity of both imprinted and nonimprinted cryogels was investigated. The maximum binding capacity for the MIP column was found to be as 79.41 mg g−1 dry cryogel, that is four times higher than the NIP column under the optimum conditions (pH 5.0, flow rate: 1.0 mL min−1, T: 25 °C). Moreover, selectivity experiments were performed by using two interference proteins as myoglobin (Mb) and cytochrome c (Cyt-c) and the relative selectivity coefficients (k') for Hb/Mb and Hb/Cyt-c pairs were determined as 36.59 and 37.22, respectively.

中文翻译：

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一种用于从血清中纯化血红蛋白的新型镧系元素螯合物分子印迹冷冻凝胶：地中海贫血诊断的替代方法

摘要 蛋白质的纯化和分析是了解其功能和与其缺乏或缺陷相关的疾病的重要手段。在这项研究中，合成了一种新的基于镧系元素螯合物的分子印迹聚合物 (MIP)，用于在存在各种干扰分子的情况下从人血清中选择性分离血红蛋白 (Hb)。根据低温聚合技术，使用复合官能单体 N-甲基丙烯酰氨基安替比林 (MAAP)-Ce(III) 和甲基丙烯酸 2-羟乙酯 (HEMA) 制备了 Hb 印迹聚合物。在没有模板 Hb 分子的情况下，非印迹冷冻凝胶 (NIP) 也在相同的聚合条件下制备。pH、初始 Hb 浓度、流速的影响，研究了温度和离子强度对印迹和非印迹冷冻凝胶的结合能力的影响。发现 MIP 柱的最大结合容量为 79.41 mg g-1 干冷冻凝胶，比最佳条件下的 NIP 柱高四倍（pH 5.0，流速：1.0 mL min-1，T：25 ℃）。此外，通过使用两种干扰蛋白作为肌红蛋白 (Mb) 和细胞色素 c (Cyt-c) 进行选择性实验，Hb/Mb 和 Hb/Cyt-c 对的相对选择性系数 (k') 确定为 36.59 和 37.22 ， 分别。