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用于免疫球蛋白G吸附的L-赖氨酸印迹冷冻凝胶的合成

Synthesis of L-lysine imprinted cryogels for immunoglobulin G adsorption.

作者信息

Çulha Senem, Armutcu Canan, Uzun Lokman, Şenel Serap, Denizli Adil

机构信息

Hacettepe University, Department of Chemistry, Beytepe, Ankara, Turkey.

Hacettepe University, Department of Chemistry, Beytepe, Ankara, Turkey.

出版信息

Mater Sci Eng C Mater Biol Appl. 2015;52:315-24. doi: 10.1016/j.msec.2015.03.059. Epub 2015 Mar 27.

DOI:10.1016/j.msec.2015.03.059
PMID:25953573
Abstract

L-Lysine imprinted poly(2-hydroxyethyl methacrylate-co-N-methacryloyl-L-aspartic acid) [P(HEMA-co-MAAsp)] cryogels were synthesized and characterized with Fourier transform infrared spectroscopy, scanning electron microscopy, surface area measurements, swelling, and squeezing tests. Specific surface area for imprinted cryogel was 34.2m(2)/g while the value was 21.3m(2)/g for non-imprinted cryogel. IgG adsorption from aqueous solution was examined in continuous mode examining the factors effecting adsorption capacity such as pH, concentration, flow rate, temperature, ionic strength, and incubation time. 0.5M NaCl was used as desorption agent. The IgG adsorption capacity was determined as 55.1 mg/g for 1.0 mg/mL IgG original concentration at 25.0°C while pH and flow rate were 7.0 and 0.5 mL/min, respectively. When human serum was used as IgG source, the removal of 90.4% of crude IgG was attained for 1/20 diluted plasma sample. The imprinted cryogel was used in ten successive cycles without significant loss in adsorption capacity. The cryogel was determined to be 1.79 times more selective to IgG than albumin and 1.45 times more selective than hemoglobin. The adsorption behavior well suited to Langmuir isotherm and the kinetics followed pseudo-second-order model. Thermodynamic parameters ΔH°, ΔS° and ΔG° for this adsorption process were also calculated.

摘要

合成了L-赖氨酸印迹聚(甲基丙烯酸2-羟乙酯-co-N-甲基丙烯酰-L-天冬氨酸)[P(HEMA-co-MAAsp)] 冷冻凝胶,并通过傅里叶变换红外光谱、扫描电子显微镜、表面积测量、溶胀和挤压试验对其进行了表征。印迹冷冻凝胶的比表面积为34.2m²/g,而非印迹冷冻凝胶的值为21.3m²/g。以连续模式研究了从水溶液中吸附IgG的情况,考察了影响吸附容量的因素,如pH值、浓度、流速、温度、离子强度和孵育时间。使用0.5M NaCl作为解吸剂。在25.0°C时,对于初始浓度为1.0mg/mL的IgG,当pH值和流速分别为7.0和0.5mL/min时,IgG的吸附容量测定为55.1mg/g。当使用人血清作为IgG来源时,对于1/20稀释的血浆样品,可去除90.4%的粗IgG。印迹冷冻凝胶连续使用十个循环,吸附容量无明显损失。测定该冷冻凝胶对IgG的选择性比对白蛋白高1.79倍,比对血红蛋白高1.45倍。吸附行为非常符合朗缪尔等温线,动力学遵循准二级模型。还计算了该吸附过程的热力学参数ΔH°、ΔS°和ΔG°。

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