溶菌酶分子印迹聚合物膜的制备及其吸附性能
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摘要
以溶菌酶为模板蛋白质,结合分子印迹技术在硅烷化的基质玻片上制备了溶菌酶分子印迹聚合物膜。实验优化了溶菌酶聚合物膜的印迹体系,考察了溶菌酶分子印迹聚合物膜的吸附平衡时间、最大吸附量、特异识别能力、重复使用性以及对实际样品中溶菌酶的分离情况。结果表明,在最优条件下,制备的分子印迹聚合物膜对溶菌酶具有特异吸附能力,印迹因子为3.0,吸附平衡时间为5 min,吸附行为符合Langmuir吸附模型,理论最大吸附量为42.5 mg/g,实际样品中的吸附量为30 mg/g。且此印迹聚合物膜在重复使用5次后,最大吸附量仅下降了5%,具有良好的重复使用性。该方法为复杂生物样品中目标蛋白质的分离富集提供了一种快速、高效的手段。
In this work, a novel polymer membrane molecularly imprinted on a silanized glass substrate was prepared. Lysozyme was chosen as the template protein, and acrylamide and N,N′-methylenebisacrylamide were used as the functional monomer and crosslinking agent, respectively. The adsorption equilibrium time, maximum adsorption capacity, specific recognition ability, reusability, and application of the prepared lysozyme-imprinted polymer membrane were investigated experimentally. The prepared polymer membrane exhibited special recognition ability and could quickly adsorb lysozyme. Under optimal conditions, the adsorption equilibrium time of the prepared polymer membrane for lysozyme was 5 min, and the adsorption behavior conformed to the Langmuir adsorption model; the theoretical maximum adsorption capacity was 42.5 mg/g, and the adsorption capacity for an egg sample was 30 mg/g. Additionally, the polymer membrane showed a high selectivity toward the template protein, lysozyme, and exhibited no obvious adsorption of several interfering proteins; the imprinting factor was 3.0. The maximum adsorption capacity of the polymer membrane decreased by 5% after being reused five times. The polymer membrane was used for the adsorption of lysozyme in samples, and good results were obtained. This strategy has potential for the separation and enrichment of target proteins in complex biological samples.
In this work, a novel polymer membrane molecularly imprinted on a silanized glass substrate was prepared. Lysozyme was chosen as the template protein, and acrylamide and N,N′-methylenebisacrylamide were used as the functional monomer and crosslinking agent, respectively. The adsorption equilibrium time, maximum adsorption capacity, specific recognition ability, reusability, and application of the prepared lysozyme-imprinted polymer membrane were investigated experimentally. The prepared polymer membrane exhibited special recognition ability and could quickly adsorb lysozyme. Under optimal conditions, the adsorption equilibrium time of the prepared polymer membrane for lysozyme was 5 min, and the adsorption behavior conformed to the Langmuir adsorption model; the theoretical maximum adsorption capacity was 42.5 mg/g, and the adsorption capacity for an egg sample was 30 mg/g. Additionally, the polymer membrane showed a high selectivity toward the template protein, lysozyme, and exhibited no obvious adsorption of several interfering proteins; the imprinting factor was 3.0. The maximum adsorption capacity of the polymer membrane decreased by 5% after being reused five times. The polymer membrane was used for the adsorption of lysozyme in samples, and good results were obtained. This strategy has potential for the separation and enrichment of target proteins in complex biological samples.
引文
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[18]Kong Q,Wang Y,Zhang L,et al.Sens Actuators B,2017,243:130
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