麦胚球蛋白分子印迹聚合物制备及表征
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摘要
运用蛋白质印迹法对麦胚球蛋白中的免疫组分进行分离提纯。以人血丙种球蛋白为模板,间氨基苯硼酸为功能单体,以活化的壳聚糖球为载体,制备对麦胚球蛋白有特异性吸附的2种印迹材料。利用傅里叶红外色谱及扫描电镜对印迹聚合物进行表征。通过静态吸附及等温吸附试验研究其吸附性能,并分析MIPs-麦胚蛋白吸附-解析附的过程。结果表明,以2种形态壳聚糖微球为载体的印迹聚合物对麦胚球蛋白组分的有效吸附量分别为20.75,3.27 mg/g;分离因数达到1.92,1.04。对印迹聚合物A建立Langmuir吸附模型,得到吸附平衡常数K为6.11。通过分析发现该材料对麦胚球蛋白免疫组分存在特异性吸附且能够有效洗脱。该聚合物的制备为进一步研究开发麦胚球蛋白及医药级球蛋白的纯化提供了新思路。
The novel MIPs were prepared via method of one-step polymerization, in which human normal immunoglobulin(Ig G) as template molecule, 3-Aminophenylboronic acid(APBA) as functional monomers, glutaraldehyde solution as crosslinker. The as-prepared polymers were characterized by fourier transform infrared(FT-IR) and scanning electron microscopy(SEM). The specific adsorption equilibrium and selectivity of one kind of MIPs were evaluated by static adsorption experiment and adsorption model. The results demonstrated that the imprinted polymer layers were successfully fabricated on the surface of CTS microspheres, resulting a better performance of adsorption capacity and specificity adsorption. The corresponding maximum adsorption capacities of WGG onto two MIPs respectively were 125 mg/g and 85.7 mg/g and their quantity of effective adsorption respectively were 20.75, 3.27 mg/g. The MIPs with high imprinting factor of 1.92, 1.04 indicated that they were of good selectivity. The adsorption equilibrium constant K was respectively 6.11 obtained through Langmuir adsorption model. This protocol will be further employ for the extraction and separation of immune components from WGG.
The novel MIPs were prepared via method of one-step polymerization, in which human normal immunoglobulin(Ig G) as template molecule, 3-Aminophenylboronic acid(APBA) as functional monomers, glutaraldehyde solution as crosslinker. The as-prepared polymers were characterized by fourier transform infrared(FT-IR) and scanning electron microscopy(SEM). The specific adsorption equilibrium and selectivity of one kind of MIPs were evaluated by static adsorption experiment and adsorption model. The results demonstrated that the imprinted polymer layers were successfully fabricated on the surface of CTS microspheres, resulting a better performance of adsorption capacity and specificity adsorption. The corresponding maximum adsorption capacities of WGG onto two MIPs respectively were 125 mg/g and 85.7 mg/g and their quantity of effective adsorption respectively were 20.75, 3.27 mg/g. The MIPs with high imprinting factor of 1.92, 1.04 indicated that they were of good selectivity. The adsorption equilibrium constant K was respectively 6.11 obtained through Langmuir adsorption model. This protocol will be further employ for the extraction and separation of immune components from WGG.
引文
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[2]BRANDOLINI A,HIDALGO A.Wheat germ:not only a by-product[J].International Journal of Food Sciences&Nutrition,2012,63(sup1):71-74.
[3]WILLIAMJ H,WILLIAMH V,CHARLENEK T,et al.Effect of high temperature on albumin and globulin accumulation in the endosperm proteome of the developing wheat grain[J].Journal of Cereal Science,2009,49(1):12-23.
[4]XU B,ZHOU S L,MIAO W J,et al.Study on the stabilization effect of continuous microwave on wheat germ[J].Journal of Food Engineering,2013,117(1):1-7.
[5]孙震,周惠明.小麦胚芽中水溶性和盐溶性提取物的免疫活性研究[J].中国粮油学报,2001,16(5):29-32.
[6]朱科学,郭晓娜,彭伟,等.采用Raman光谱研究小麦胚芽8S球蛋白的二级结构[J].食品科学,2009,30(19):32-34.
[7]MOHAJERI S A,EBRAHIMI S A.Preparation and characterization of a lamotrigine imprinted polymer and its application for drug assay in human serum[J].J Sep Sci,2008,31(20):3595-3602.
[8]WANG H,LI W,HE X,et al.m-Aminophenylboronic acid as a functional monomer for fabricating molecularly imprinted polymer for the recognition of bovine serum albumin[J].Reactive&Functional Polymers,2008,68(9):1291.
[9]NICOLE M,NICHOLAS A.Molecularly imprinted polymers with specific recognition for macromolecules and proteins[J].Science Direct,2008,33(19):271-288.
[10]高婷,郭敏杰,樊志,等.牛血清白蛋白分子印迹壳聚糖树脂的制备[J].天津工业大学学报,2010,25(1):20-23.
[11]刁大鹏,黄继红,李锦,等.小麦胚芽球蛋白提取工艺及分子质量测定[J].河南工业大学学报(自然科学版),2013,34(1):52-53.
[12]马豫峰,蔡继业.壳聚糖与牛血清白蛋白分子印迹聚合物的制备与表征[J].高分子材料科学与工程,2007,23(2):235-237,241.