基于分子印迹聚合物的SPR传感器用于牛血清蛋白的检测

Molecularly Imprinted Polymers for Determination of Bovine Serum Albumin Based Surface Plasmon Resonance Sensor

以牛血清蛋白(BSA)为模板分子、多巴胺为功能单体和交联剂、高碘酸钠为氧化剂,制备了BSA表面印迹表面等离子共振(SPR)生物传感器。通过聚丙烯酸(PAA)与BSA的氢键作用预先固定BSA,增加印迹效率,同时在聚多巴胺表面非印迹区域修饰部分水解的聚(2-甲基-2-噁唑啉)(PMOXA-EI)抵抗蛋白质的非特异性吸附。通过X射线光电子能谱、原子力显微镜、可变角光谱椭偏仪和静态水接触角对制备的BSA表面印迹SPR生物传感器进行表征。对质量浓度为0.1~10μg/mL的BSA水溶液进行SPR吸附研究,检测限和定量限分别达到了53 ng/mL和161 ng/mL。以β-乳球蛋白、卵白蛋白、溶菌酶和细胞色素C为参比蛋白进行选择性研究,相应的选择性系数分别达到了4.43、3.45、3.17和3.64。上述5种蛋白混合溶液中BSA的检测回收率为97.5%~102.5%。

Bovine serum albumin(BSA)molecularly imprinted polymer(MIP)surface plasmon resonance(SPR)biosensors were prepared using BSA as the model,dopamine as the functional monomer and cross-linker and sodium periodate as the oxidant.BSA molecules were fixed in advance through hydrogen bond via introducing poly(acrylic acid)(PAA)to increase recognition sites,and partially hydrolyzed poly(2-methyl-2-oxazoline)(PMOXA-EI)was also introduced into the noncavity regions of polydopamine to resist nonspecific adsorption of proteins.The prepared BSA-MIP SPR sensor was characterized by X-ray photoelectron spectroscopy,atomic force microscopy,variable angle spectroscopic ellipsometry,and static water contact angle.SPR adsorption studies were carried out in aqueous BSA solutions with the mass concentration of 0.1—10μg/mL.The limit of detection and the limit of quantification values were obtained as 53 ng/mL and 161 ng/mL,respectively.Selectivity studies were performed againstβ-lactoglobulin,ovalbumin,lysozyme,and cytochrome C,and the corresponding selectivity coefficients were determined to be 4.43,3.45,3.17 and 3.64,respectively.Finally,BSA-MIP biosensor was used to detect BSA in the mixture solution of the above five proteins.The BSA recovery rate was 97.5%-102.5%.