基于聚硫堇和纳米银固定酶的葡萄糖生物传感器

用循环伏安法将电子媒介体硫堇电聚合在玻碳电极表面上,使其表面形成均匀的带负电的聚硫堇膜,通过静电吸附作用吸附表面带正电荷的纳米银溶胶,接着通过静电吸附带负电的葡萄糖氧化酶,最后用聚硫堇包埋电极,从而制得性能优良的葡萄糖氧化酶(GOD)生物传感器。实验发现传感器氧化峰电流与葡萄糖的浓度在1.0×10-8~5.0×10-6mol/L(r=0.9963)范围内呈良好线性关系,检出限为5.0×10-9mol/L(S/N=3)。     

基于多壁碳纳米管-硫堇/Nafion纳米复合物修饰的人IgG免疫传感器的研究

制备了基于多壁碳纳米管-硫堇/Nafion纳米复合物的人IgG免疫传感器.阻抗谱、循环伏安研究表明,该免疫传感器对人IgG的检测具有优异的性能,其对人免疫球蛋白G(IgG)浓度的定量测定线性范围为1.0 ～ 200μg/L,检出限为0.25 μg/L,线性相关系数R=0.9950.该免疫传感器可用于对人血清中IgG的检...     

基于HGB/PTh/AuNCs无标记癌胚抗原免疫传感器的研究

本文研制了一种基于中空石墨烯球(HGB)/聚硫堇/金纳米笼的无标记型癌胚抗原传感器。将合成的中空石墨烯球修饰于玻碳电极(GCE)表面,在其上电聚合硫堇形成多孔聚硫堇(PTh)复合膜后吸附固定金纳米笼,然后将癌胚抗体(anti-CEA)固定到电极表面,制得无标记型癌胚抗原(CEA)免疫传感器。当抗体与抗原发生免疫反应时,形成的复合物阻碍了电子传递,从而降低聚硫堇的电催化活性。通过示差脉冲伏安法(DPV)检测聚硫堇的响应电流信号的降低,实现对CEA的无标记检测。在最优的实验条件下,检测癌胚抗原的线性范围为0.5~80 ng·mL-1,线性相关系数为0.9932,检测限为0.17 ng·mL-1。该免疫传感器可用于临床上CEA的检测。在血清样品中进行了CEA的检测,结果令人满意。     

DNA电化学生物传感器测定水中痕量铅

采用物理涂附法将单链DNA固定到金表面得到ssDNA/Au修饰电极。研究表明,该电极在pH 5,0.2 mol/L NaAc-HAc缓冲底液中,可用于测定水中痕量Pb2 。Pb2 以Pb-DNA络合物的形式吸附在电极上,以示差脉冲伏安法测定Pb2 ,在0.15 V(vsSCE)处有灵敏的氧化峰,其峰电流与Pb2 浓度在5.0×10-8~1.0×10-6mol/L范围内呈良好的线性关系,此法的检测限为2.0×10-8mol/L,该修饰电极的稳定性和抗干扰性都较好。用该电极对饮用水中的Pb进行检测,得到了满意的结果。     

石墨烯-壳聚糖修饰电极构建石杉碱甲免疫传感器

通过将石杉碱甲抗体(anti-HupA)固定到由还原态氧化石墨烯/壳聚糖(r GO/CS)复合膜修饰的玻碳电极表面,制备高灵敏度的电化学免疫传感器,利用石杉碱甲抗原抗体特异性反应可阻断[Fe(CN)6]3-/4-氧化还原反应体系电子转移的特点,建立了检测石杉碱甲(Hup A)的电化学免疫传感器分析方法。采用循环伏安法(CV)、交流阻抗法(EIS)及差示脉冲伏安法(DPV)研究Hup A在[Fe(CN)6]3-/4-氧化还原体系的电化学行为,利用DPV法测定Hup A浓度。在优化条件下,15 mmol/L K3[Fe(CN)6]+0. 1 mol/L KCl+0. 1 mol/L PBS(pH 6. 5)测试体系中,Hup A浓度的对数与峰电流差值在1. 0×10-13~1. 0×10-10mol/L范围内呈良好的线性关系,相关系数(r)为1. 000 0,检出限为3. 0×10-14mol/L,回收率为99. 4%~102%。该方法具有特异性强、灵敏度高、操作简单、绿色环保等特点,可用于中药材中石杉碱甲浓度的快速检测。     

基于聚2,6-二氨基吡啶膜及纳米金修饰的癌胚抗原免疫传感器研究

在玻碳电极表面电聚合2,6-二氨基吡啶(pPA), 利用硫堇(Thi)、纳米金(nano-Au)固载癌胚抗体, 制得稳定性好、灵敏度较高、线性范围宽的电流型免疫传感器. 通过循环伏安法考察了该免疫传感器的电化学特性, 在优化的实验条件下, 该免疫传感器的峰电流随着检测溶液中癌胚抗原(CEA)浓度的增大而减小, 并在0.5～20和20～160 ng/mL CEA范围内呈现出良好的线性关系, 检测下限为0.2 ng/mL. 该免疫传感器具有制作简单、重现性好、线性范围宽等优点, 可用于临床上对CEA的检测.     

基于纳米银与有机多孔材料/纳米金修饰的甲胎蛋白免疫传感器研究

在金电极表面电沉积银为氧化还原探针,利用有机多孔材料(PTC-NH2)、纳米金(nano-Au)固载甲胎蛋白抗体(anti-AFP),制备出用于检测甲胎蛋白(AFP)的安培型免疫传感器。通过交流阻抗技术、循环伏安法研究了电极的电化学特性,考察了孵育时间、测试液pH值等实验条件对传感器性能的影响,并利用扫描电子显微镜(SEM)对电极的修饰过程进行了表征。该传感器对AFP有良好的电流响应,线性范围分别为1.0～20.0ng/mL和20.0～60.0 ng/mL,检测限为0.6 ng/mL。     

基于纳米金/硫堇层层自组装的新型电流型酶-癌胚抗原免疫传感器

将Nafion 膜固定在金电极(Au)表面, 通过静电吸附和共价键合作用将硫堇(Thi)和纳米金颗粒(nano-Au)层层自组装到Nafion膜修饰的金电极表面. 再通过形成的纳米金单层吸附癌胚抗体(anti-CEA), 最后用辣根过氧化物酶(HRP)代替牛血清白蛋白(BSA)封闭电极上的非特异性吸附位点, 并同时起到放大响应电流信号的作用, 从而制得高灵敏、高稳定电流型酶-癌胚抗原(CEA)免疫传感器. 通过循环伏安和交流阻抗考察了电极表面的电化学特性, 并对该免疫传感器的性能进行了详细的研究. 该传感器对CEA检测的线性范围为2.5~80.0 ng/mL, 检测限为0.90 ng/mL.     

基于石墨烯-甲苯胺蓝复合物癌胚抗原电化学免疫传感器

研究了在PBS缓冲介质中,一种检测癌胚抗原的新型免标记电化学免疫传感器的制备及应用,石墨烯与甲苯胺蓝复合物饰于玻碳电极表面,通过循环伏安法对修饰的电极进行表征.基于以[Fe(CN)6]3-/4-为氧化还原探针,癌胚抗原抗体反应引起[ Fe(CN)6] 3-/4-探针的电流响应的变化,来实现癌胚抗原的检测,癌胚抗原的浓度...     

基于聚硫堇和层层组装碳纳米管/HRP的过氧化氢传感器研究

将电聚合和层层组装方法有效结合构筑了聚硫堇（PTH）电子介体修饰的碳纳米管（CNTs）/辣根过氧化物酶（HRP）多层膜无试剂H2O2传感器.利用电化学阻抗谱对CNTs/HRP多层膜的组装过程进行监测,用循环伏安法和计时电流法研究了该HRP电极的电化学行为.探讨了酶组装层数、工作电位、pH值和碳纳米管对电极响应的影响.该...     

基于纳米金与碳纳米管-纳米铂-壳聚糖纳米复合物固定癌胚抗原免疫传感器的研究

采用纳米金(nano-Au)、多壁碳纳米管-纳米铂-壳聚糖的纳米复合物(MWNT-Pt-CS)及电子媒介体硫堇(Th)固载抗体制得高灵敏癌胚抗原免疫传感器．首先, 于壳聚糖溶液中用NaBH4还原H2PtCl6, 并将多壁碳纳米管分散于其中制得碳纳米管-纳米铂-壳聚糖纳米复合物, 并将其滴涂在玻碳电极上成膜; 然后, 吸附电子媒介体硫堇制得硫堇/碳纳米管-纳米铂-壳聚糖(Th/MWNT-Pt-CS)修饰电极．利用壳聚糖和硫堇分子中大量的氨基固定纳米金并吸附癌胚抗体(anti-CEA); 最后, 用辣根过氧化物酶(HRP)封闭活性位点从而制得高灵敏电流型免疫传感器．在优化的实验条件下, 该传感器响应的峰电流值与癌胚抗原(carcinoembryonic antigen)浓度在0.5～10和10～120 ng/mL的范围内保持良好的线性关系, 检测限为0.2 ng/mL．     

Disposable Amperometric Label-Free Immunosensor on Chitosan–Graphene-Modified Patterned ITO Electrodes for Prostate Specific Antigen

A facile and highly sensitive determination of prostate-specific antigen (PSA) is of great significance for the early diagnosis, monitoring and prognosis of prostate cancer. In this work, a disposable and label-free electrochemical immunosensing platform was demonstrated based on chitosan–graphene-modified indium tin oxide (ITO) electrode, which enables sensitive amperometric determination of PSA. Chitosan (CS) modified reduced graphene oxide (rGO) nanocomposite (CS–rGO) was easily synthesized by the chemical reduction of graphene oxide (GO) using CS as a dispersant and biofunctionalizing agent. When CS–rGO was modified on the patterned ITO, CS offered high biocompatibility and reactive groups for the immobilization of recognition antibodies and rGO acted as a transduction element and enhancer to improve the electronic conductivity and stability of the CS–rGO composite film. The affinity-based biosensing interface was constructed by covalent immobilization of a specific polyclonal anti-PSA antibody (Ab) on the amino-enriched electrode surface via a facile glutaraldehyde (GA) cross-linking method, which was followed by the use of bovine serum albumin to block the non-specific sites. The immunosensor allowed the detection of PSA in a wide range from 1 to 5 ng mL⁻¹ with a low limit of detection of 0.8 pg mL⁻¹. This sensor also exhibited high selectivity, reproducibility, and good storage stability. The application of the prepared immunosensor was successfully validated by measuring PSA in spiked human serum samples.     

新型有机-无机氧化还原复合膜层层组装的无试剂高灵敏电流型前列腺特异性抗原免疫传感器研究

以前列腺特异性抗原(PSA)和前列腺特异性抗体(anti-PSA)为生物模型分子, 采用电沉积技术和共价键合作用, 研制了新型高灵敏电流型免疫传感器. 利用具有良好导电性和热稳定性的新型有机材料[苝四甲酸二酐(PTCDA)衍生物, 简写为PTC-NH2]膜具有的多孔结构, 该膜可与电沉积制得的冰晶状普鲁士蓝(PB)颗粒进行层层组装镶嵌, 形成多层稳定的有机-无机氧化还原复合膜以增加 PB 的固定量和稳定性, 从而提高电极的电流响应信号; 同时, 通过复合膜表面丰富的氨基吸附大量纳米金以增加抗体的固定量, 从而提高免疫传感器的灵敏度. 利用扫描电子显微镜(SEM)和X射线光电子能谱仪(XPS)对PTC-NH2膜的形貌和结构进行表征, 通过循环伏安法考察了电极修饰过程的电化学特性, 详细研究了该免疫传感器的性能. 该免疫电极对前列腺特异性抗原检测的线性范围为0.5～16.0 ng/mL, 相关系数为0.985, 检测限为0.02 ng/mL. 实验结果表明, 利用该方法制备的免疫传感器具有灵敏度高、稳定性和选择性好等优点.     

磁性纳米金共价固定癌胚抗原单克隆抗体的电流型免疫传感器

合成了Fe3O4/Au磁性复合纳米粒子, 在粒子表面通过自组装硫脲分子使表面氨基化, 再用戊二醛共价交联固定癌胚抗原抗体(anti-CEA). 在外加磁场的作用下, 将anti-CEA复合磁性粒子吸附在固体石蜡碳糊电极表面, 制成了新型电流型免疫传感器. 免疫电极在含有癌胚抗原CEA和辣根过氧化物酶标记的癌胚抗原(HRP-CEA)的混合溶液中温育, CEA和HRP-CEA与固定在电极表面的anti-CEA发生竞争反应, 导致HRP对H2O2的催化降解作用的改变, 从而可间接测定CEA. 由于标记的HRP可催化降解H2O2, 导致媒介体间苯二酚浓度改变, 使测定的灵敏度大大提高. 响应电流与CEA质量浓度的对数在2~160 ng/mL的范围内呈线性关系, 检出限为0.57 ng/mL(3σ法). 该免疫传感器具有制作简单、价廉及表面易于更新等特点.     

基于透明质酸及硫堇层层自组装的前列腺特异性抗原免疫传感器的研究

将带负电荷的透明质酸(HA)和带正电荷的电子媒介体硫堇(Thi)层层自组装到壳聚糖修饰的玻碳电极表面,利用硫堇上的氨基固定纳米金(nano-Au)并固载前列腺特异性抗体(anti-PSA),制得新型前列腺特异性抗原电流型免疫传感器。通过循环伏安法(CV)、微分脉冲伏安法(DPV)考察了该免疫传感器的电化学特性,并对影响该免疫传感器性能的各种因素进行详细研究。在优化实验条件下,此免疫传感器的线性范围为0.5~4μg/L和4~25μg/L,检出限为0.2μg/L。该免疫传感器具有制备简单、灵敏度高、稳定性好等优点。     

Microcystin-(Leucine-Arginine) Immunosensor Based on Iron(II,III) Magnetic Nanoparticles

An electrochemical immunosensor for microcystin-(leucine-arginine) based on magnetic bionanoparticles and iron(II, III) oxide was developed. The bionanoparticles were prepared by cross-linking antibodies of microcystin-(leucine-arginine) and amino-functionalized magnetic iron(II, III) oxide nanoparticles with glutaraldehyde, followed by immobilization on the surface of a magnetic electrode. The immunosensor was based on the model of direct competition, as microcystin-(leucine-arginine) and horseradish peroxidase-conjugated microcystin-(leucine-arginine) competitively combined with immobilizing antibodies. The peak current of differential pulse voltammetry (DPV) decreased with an increase of microcystin-(leucine-arginine) concentration after antigen-antibody reaction. When the background current was stabilized, the anodic peak current response and change in peak response were recorded. Under the optimized conditions, the change in response was proportional to the microcystin-(leucine-arginine) concentration between 0.010 and 100 µg/L with a limit of detection equal to 0.009 µg/L. Amperometry was adopted to determine microcystin-(leucine-arginine); the linear dynamic range was 0.10 to 100 µg/L with a detection limit of 0.08 µg/L. The method was successfully applied in the determination of microcystin-(leucine-arginine) in river water and the recoveries were between 90.2% and 110.5%.     

基于复合纳米微粒修饰电极的氯霉素快速检测用磁场可控一次性安培免疫传感器研究

在丝网印刷碳电极(SPCE)表面涂敷一层碳纳米管(CNT)/二茂铁(Fc)/Nafion膜,进而通过外磁场作用将包被了氯霉素-牛血清蛋白(CAP-BSA)的纳米金磁微粒[Fe3O4(核)/Au(壳),简称GMP]吸附到其表面,构建了可用于CAP检测的磁场可控一次性安培免疫传感器.采用扫描电镜(SEM)、X射线荧光光谱(XRFS)表征了免疫传感器的制备过程,采用循环伏安法(CV)和示差脉冲伏安法(DPV)研究了免疫传感器的电化学性质,结合竞争性免疫分析法测定了CAP浓度.免疫传感器在含有100ng·mL-1anti-CAP的25μLpH=6.0磷酸盐缓冲溶液(PBS)中加入不同浓度的CAP,并在25℃下温育6min,DPV还原峰电流上升百分比(CI%)与CAP浓度在0.2～80.0ng·mL―1范围内呈线性关系,检测限为0.11ng·mL―1.用于牛奶样品中CAP检测并和标准HPLC方法对照,结果一致,回收率在88%～104%之间.该免疫传感器灵敏快速、外磁场可控、样品用量小、可抛弃,有望用于食品中痕量CAP快速检测.     

纳米粒子标记激光诱导荧光免疫分析测定癌胚抗原

建立了一种核壳型SiO₂荧光纳米粒子标记的激光诱导荧光免疫测定的新方法. 该方法用 掺杂的核壳型SiO₂荧光纳米粒子为标记物, 采用双抗体夹心法, 在载玻片上固定鼠抗人癌胚抗原(CEA)单克隆抗体, 加入1～2 μL样品, 再加入纳米粒子标记的另一种鼠抗人癌胚抗原(CEA)单克隆抗体. 识别后在光导纤维激光诱导荧光毫米阵列检测平台上测量荧光信号, 进行定量. 在优化条件下, 荧光强度和癌胚抗原的量在1～80 pg的范围内具有良好的线性关系, 相关系数r＝0.9933, 方法的检出限为0.3 pg (20 amol). 对40 pg的CEA平行测定5次, 相对标准偏差为5.5%.     

Electroanalytical Determination of Carcinoembryonic Antigen at a Silica Nanoparticles/Titania Sol–Gel Composite Membrane‐Modified Gold Electrode

A highly hydrophilic and nontoxic colloidal silica nanoparticle/titania sol–gel composite membrane was prepared on a gold electrode via a chemical vapor deposition method. With carcinoembryonic antigen (CEA) as a model antigen and encapsulation of carcinoembryonic antibody (anti‐CEA) in the composite architecture, this membrane could be used for reagentless electrochemical immunoassay. The presence of silica nanoparticles provided a congenial microenvironment for adsorbed biomolecules. The formation of immunoconjugate by a simple one‐step immunoreaction between CEA in sample solution and the immobilized anti‐CEA introduced the change in the potential. The modified procedure was further characterized by electrochemical impedance spectroscopy and cyclic voltammetry. Compared to the commonly applied methods, i.e., the TiO₂ direct embedding procedure, this strategy could allow for antibodies immobilized with higher loading amount and better retained immunoactivity. The resulting immunosensor exhibited high sensitivity, good precision, acceptable stability, accuracy, reproducibility and wide linear range from 1.5 to 240 ng mL^?1 with a detection limit of 0.5 ng mL^?1 at 3σ. Analytical results of clinical samples show that the developed immunoassay is comparable with the enzyme‐linked immunosorbent assays (ELISAs) method, implying a promising alternative approach for detecting CEA in the clinical diagnosis. Furthermore, this composite membrane could be used efficiently for the entrapment of other biomarkers and clinical applications.