糖蛋白印迹二维光子晶体水凝胶传感器的构建

制备了一种特异性识别糖蛋白的二维光子晶体分子印迹水凝胶。以刀豆蛋白A(Con A)为模板分子,甲叉双丙烯酰胺(MBA)为交联剂,丙烯酰胺(AAm)与4-乙烯基苯硼酸为功能单体,N,N,N′,N′-四甲基乙二胺(TEMED)为引发剂,过硫酸铵(APS)为催化剂,在聚苯乙烯二维光子晶体(PC)表面产生聚合反应形成水凝胶,接着用10%的乙酸溶液洗脱模板分子,制备了可对Con A特异性识别的二维光子晶体分子印迹水凝胶(MIPs)传感器。结果表明,当MIPs的交联度为10%时,德拜环的直径变化最显著,对目标物的识别最灵敏。该传感器对Con A在0~0.5 mg/mL范围内具有良好的响应与识别能力,可用于目标物的富集和即时检测。同时,苯硼酸的引入,增加了印迹空穴的识别位点,进一步改善了印迹材料对糖类分子的特异识别性。该水凝胶传感器在卵清白蛋白(OVA)、牛血红蛋白(BHb)、溶菌酶(Lyz)、胰蛋白酶(Try)溶液中德拜环的直径变化远小于识别相同浓度的Con A引起的德拜环直径的变化,表现出良好的选择性。该方法操作简单,成本低廉,可重复使用,不需特殊仪器设备,可拓展到其它糖蛋白,实现糖类目标蛋白的即时快速检测。

Construction of a Two dimensional Photonic Crystal Hydrogel Sensor with Glycoproteins Imprinting

A two-dimensional photonic crystal molecularly imprinted hydrogel(MIPs) sensor with specific recognition to glycoproteins was prepared by using concanavalin A(Con A) as the imprinted molecule,methylidenebisacrylamide(MBA) as the cross-linking agents,acrylamide(AAm) and 4-vinylbenzeneboronic acid as the functional monomers,tetramethylethylenediamine(TEMED) as an initiator and ammonium persulfate(APS) as a catalyst.Polymerization was carried out to form a Con A imprinted hydrogel on the surface of two-dimensional photonic crystal.After elution with 10% acetic acid solution,a MIPs sensor for specific recognition to Con A was constructed.Results showed that the diameter of Debye ring changed most significantly,and the target object was the most sensitive to be identified when the cross-linking degree of MIPs was 10%.The sensor could be used in the enrichment and real-time detection of target molecules as it had a good response to Con A in the concentration range of 0-0.5 mg/mL.Moreover,the introduction of phenyl boronic acid increased the recognition sites of the imprinted cavities,and further improved the specific recognition of the MIPs toward the carbohydrate molecules.The hydrogel sensor presented a good selectivity when binding to the proteins in ovalbumin(OVA),bovine hemoglobin(BHb),lysozyme(Lyz),trypsin(Try) solution under the same concentration.This method is easy to operate,low cost,reusable and needless for any special equipments.Furthermore,it could be extended to the real-time and fast detection for other glycoproteins.