邻苯二胺—过氧化氢—辣根过氧化物酶酶联免疫示差脉冲伏安法测定人血清…

建立了邻苯二胺（ＯＰＤ）－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）酶联示差脉冲伏安分析体系并用于测定人血清中类风湿因子（ＲＦ），ＨＲＰ催化Ｈ２Ｏ２氧化ＯＰＤ所形成麦催化产物在ＰＨ２．０磷酸盐－枸椽酸缓冲溶液中于－０．１８Ｖ左右产生－灵敏示差脉冲伏安峰，在ＲＦ浓度在１．２５－２０．０Ｕ／ｍＬ之间与峰电流呈线性关系，应用此峰检测人血清ＲＦ的检测限低至０．２８Ｕ／ｍＬ。该法较相同条件下ＥＬＩＳＡ以光度测定法的     

联苯胺-H_2O_2-HRP伏安酶联免疫分析体系测定植物病毒TMV和TRSV

提出了联苯胺－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）伏安酶联免疫分析体系测定植物病毒烟草花叶病毒（ＴＭＶ）和烟草环斑病毒（ＴＲＳＶ）的新方法。ＨＲＰ标记的羊抗兔酶标记抗体ＩｇＧ－ＨＲＰ可以催化Ｈ２Ｏ２氧化联苯胺的反应,其氧化产物在Ｂｒｉｔｏｎ－Ｒｏｂｉｎｓｏｎ（ＢＲ）缓冲溶液中在－０．６２Ｖ（ＳＣＥ）左右产生灵敏的线性扫描二阶导数伏安峰,可以测定ＩｇＧ－ＨＲＰ。根据ＩｇＧ－ＨＲＰ与植物病毒及其抗血清的免疫反应,可以间接测定植物病毒。本法测定ＴＭＶ的检出限为０．２５ｎｇ／ｍＬ,线性范围为０．２５～５０００ｎｇ／ｍＬ;测定ＴＲＳＶ的检出限为１．５ｎｇ／ｍＬ,线性范围为１．５～３０００ｎｇ／ｍＬ;测定ＴＭＶ烟草病叶澄清液的最高稀释比为１∶１００００。检测灵敏度高于酶联免疫吸附显色光度法（ＥＬＩＳＡ）     

OAP-H2O2-HRP伏安酶联免疫分析新体系测定人血清总甲状腺素

提出邻氨基酚（ＯＡＰ）－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）发酶联免疫分析新体系并用于人血清中总甲状腺素的测定。本方法以线性扫描二阶导数伏安法检测ＨＲＰ催化Ｈ２Ｏ２氧化ＯＡＰ的产物,用于游离ＨＲＰ和ＨＲＰ标记物的测定,灵敏度的高于经典的ＥＬＩＳＡ显色光度法。测定游离ＨＲＰ的线性范围为１．０×１０＾１２￣４．０×１０＾－９ｇ／ｍＬ,检测限达６．０×１０＾１３ｇ／ｍＬ。本法对总甲状腺素测定的线性范围为１     

邻氨基酚-过氧化氢-辣根过氧化物酶伏安酶联免疫分析法测定人血清甲胎蛋白

提出了邻氨基酚（ＯＡＰ）－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）伏安酶联免疫分析法测定人血清甲胎蛋白（α－ＦＰ）的新方法．该方法是将ＨＲＰ催化Ｈ２Ｏ２氧化邻氨基酸的酶催化反应与邻氨基酚的氧化产物在滴汞电极上的还原反应相偶合，在ＢＲ缓冲溶液中，在－０．４３ Ｖ（ｖｓ．ＳＣＥ）左右产生灵敏的极谱波．根据测定标记在甲胎蛋白抗体上的ＨＲＰ的量，求得发生免疫反应的 α－ＦＰ的含量。该方法对甲胎蛋白测定的线性范围为 １． ２５～４００ ｍｇ／Ｌ。用所建立的方法对病人血清样品进行了测定，并与酶联免疫吸附测定光度法（ＥＬＩＳＡ）进行对照，二者相关性很好。     

以氟苯酚—H2O2—HRP体系酶联荧光免疫分析研究：Ⅲ.F^—Al酸性铬蓝…

建立了一种检测人血清中乙肝Ｅ抗原的新荧光光度法，通过酶促反应，对氟苯酚＋Ｈ２Ｏ２→＾ＨＲＰ苯酚＋Ｆ＾－＋Ｈ２Ｏ与Ａｌ－酸性铬蓝Ｋ荧光体系相偶合，测定辣根过氧化物酶（ＨＲＰ）及其标记物，测定ＨＲＰ的线性范围为０．１９～３１ｍＵ／ｍＬ，检出限为０．０４ｍＵ／ｍＬ。     

对氟苯酚-H_2O_2-HRP体系酶联荧光免疫分析研究ⅢF~--Al-酸性铬蓝K荧光熄灭法测定人血清中乙肝E抗原

建立了一种检测人血清中乙肝Ｅ抗原的新荧光光度法．通过酶促反应：对氟苯酚＋Ｈ２Ｏ２ＨＲＰ→苯酚＋Ｆ－＋Ｈ２Ｏ与Ａｌ－酸性铬蓝Ｋ荧光体系相偶合，测定辣根过氧化物酶（ＨＲＰ）及其标记物．测定ＨＲＰ的线性范围为０．１９～３１ｍＵ／ｍＬ，检出限为０．０４ｍＵ／ｍＬ．     

OARP—H2O2—HRP伏安酶联免疫分析新体系测定人血清甲胎蛋白

本文提出邻氨基酚（ＯＡＰ－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）伏安酶联免疫分析体系并用于人血清中甲胎蛋白（αＦＰ）的测定。该方法是将ＨＲＰ催化Ｈ２Ｏ２氧化ＯＡＰ的酶催化反应与邻氨基酚的氧化中间产物（邻苯醌亚胺）在滴汞电极上的还原反应相偶合,在ＢＲ缓冲溶液中,在－０．８７Ｖ（ｖｓ,ＳＣＥ）左右产生灵敏的极谱波。根据测定标记在甲胎蛋白抗体上的ＨＲＰ的量,求得发生免疫反应的αＦＰ的含量。这该方法对甲胎蛋白测定的线性范围为１．２５－４００ｍｇ／Ｌ。用所建立的方法对病人的血清样品进行了测定,并与酶联免疫吸附测定光度法（ＥＬＩＳＡ）进行对照,二者相关性很好。     

联本胺—H2O2—HRP伏安酶联免疫分析体系测定植物病毒TMV和TRSV

提出了联苯胺－Ｈ２Ｏ２－辣根过氧化物酶伏安酶联免疫分析体系测定植株病毒烟草花叶病毒（ＴＭＶ）和烟草环斑病毒（ＴＲＳＶ）的新方法。ＨＲＰ标记的间抗兔酶标记抗体ＩｇＧ－ＨＲＰ可以催化Ｈ２Ｏ２氧化联苯胺的反应，其氧化产物在Ｂｒｉｔｔｏｎ－Ｒｏｂｉｎｓｏｎ（ＢＲ）缓冲溶液中在－０．６２Ｖ（ＳＣＥ）左右产生灵敏的线性扫描二阶导数伏安峰，可以测定ＩｇＧ－ＨＲＰ。根据ＩｇＧ－ＨＲＰ与植物病毒及其抗血清的免疫反应     

间氨基酚—过氧化氢—辣根过氧化物酶伏安酶联免疫分析新体系测定人血 …

提出间氨基酚（ＭＡＰ）－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）伏安酶联免疫分析新体系并用于人血清中总甲状腺素（Ｔ４）的测定。本方法以线性扫描二阶导数伏安法检测ＨＲＰ催化Ｈ２Ｏ２氧化ＭＡＰ的产物,用于游离ＨＲＰ和ＨＲＰ标记物的测定,灵敏度均坑于经典的ＥＬＩＳＡ显色光度法。本法对总甲状腺素测定的线性范围为０．５－３２０ｍｇ／Ｌ。用所建立的方法对人血清样品进行了测定,并与现行的ＥＬＩＳＡ显色光度法对照,二者     

脱氧核糖核酸变性和损伤的吸附伏安法研究

本文用汞电极（ＨＭＤＥ）二次导数阴极吸附伏安（ＳＤ－ＡｄＣＳＶ）和碳电极（ＧＣＥ、ＣＰＥ）导数循环伏安（ＦＤ－ＣＶ）法研究了核酸受热、紫外线、超声波和丝裂霉素Ｃ（ＭＭＣ）作用下的变性作用。在０．１ｍｏｌ／Ｌ（Ｋ２ＨＰＯ４＋ＫＨ２ＰＯ４）－０．１ｍｏｌ．Ｌ ＮａＣｌ（ｐＨ７．０）底液中，吸附的单股（ｓｓ－）和双螺旋（ｄｓ－）ＤＮＡ分别在ＨＭＤＥ上得到特征还原峰Ｐ３和Ｐ２，和在碳电极上得到氧化峰Ａ。物     

Investigation of voltammetric enzyme-linked immunoassay based on a new system of HAP-H2O2-HRP

By introducing heterocyclic compound to immunoassay system as an electrochemical substrate for the fist time, a new voltammetric enzyme-linked immunoassay system of 3-hydroxyl-2-aminopyridine (HAP)-H(2)O(2)-horseradish peroxidase (HRP) has been developed. HAP was oxidized with H(2)O(2) catalyzed by HRP, and the resulting electroactive product produced a sensitive voltammetric peak at potential of -0.36 V (vs. SCE) in Britton-Robinson (BR) buffer solution. The process of the enzyme-catalyzed reaction and the electro-reduction of the product have been investigated in detail. The linear range for detection of free HRP was from 4.0x10(-13) to 1.0x10(-9) g/mL with a detection limit of 1.2x10(-13) g/mL. The new system has been successfully applied for the assay of alpha-fetoprotein (alphaFP) in human serum ranging from 0.1 to 200 ng/mL with a detection limit of 0.1 ng/mL, which was 10 times lower than that of traditional spectrophotometric enzyme-linked immunosorbent assay (ELISA) method. HAP-H(2)O(2)-HRP voltammetric enzyme-linked immunoassay showed a promising alternative approach in the detection of alphaFP in clinical diagnosis.     

3,3',5,5'-四甲基联苯胺-H2O2-辣根过氧化物酶伏安酶联免疫分析法测定人血清免疫球蛋白E

近年来,电化学免疫分析法由于成功地将免疫反应的高选择性和电化学测定的高灵敏度相结合而越来越受到人们的重视[1]．酶联电化学免疫分析法的测定灵敏度与放射免疫法相近而又不必使用放射性同位素[2],充分显示了该法在临床检验中的优越性和发展前景．本文利用HRP催化TMB－H2O2的反应,以金电极为工作电极,用示差脉冲伏安法检测酶催化产物,建立了TMB－H2O2－HRP酶联免疫示差脉冲伏安分析体系,并成功地用于人血清IgE的测定。实验表明,本法较ELISA显色光度测定法的灵敏度高4倍,且具有更宽的线性范围,样品溶液基体对测定不产…     

Sensitive detection of tumor marker CA15-3 in human serum by capillary electrophoretic immunoassay with chemiluminescence detection

A new and sensitive non-competitive immunoassay (IA) for tumor marker carbohydrate antigen 15-3 (CA15-3) by CE coupling with ECL detection has been developed. This method is based on luminol-H(2)O(2 )reaction catalyzed by horseradish peroxidase (HRP). The optimum CE separation and CL detection conditions were investigated. After the non-competitive immunoreaction, the free HRP-labeled CA15-3 antibody (Ab*) and the bound Ab*-antigen (Ab*-Ag) complex were separated in a separation capillary and then catalyzed the CL reaction of luminol and H(2)O(2 )in a reaction capillary following the separation capillary. The calibration curve based on the peak areas of Ab*-Ag complex plotted against the concentrations of CA15-3 is in the range of 0-250 U/mL with a correlation coefficient of 0.9983 and the detection limit is 0.035 U/mL (S/N = 3). The response for five consecutive injections of 125 U/mL CA15-3 resulted in RSDs of 0.83% and 3.1% for the migration time and the peak area, respectively. The method was successfully used for the quantification of CA15-3 in human sera obtained from healthy persons and from patients with breast cancer.     

OT-H2O2-HRP伏安酶联免疫分析新体系

本文首次提出了邻联甲苯胺(OT)-H2O2-辣根过氧化物酶(HRP)伏安酶联免疫分析新体系。本方法以线性扫描二阶导数伏安法检测HRP催化H2O2氧化OT的产物, 用于游离HRP和各种HRP标记物测定, 灵敏度比经典的ELISA光度法分别高两个至四个数量级。测定游离HRP的检测限达到1.8×10^-^1^2 g/mL, 线性范围为5.0×10^-^1^2-1.0×10^-^8 g/mL。对此伏安酶联免疫分析新体系的偶合反应机理及电极还原过程也进行了详细的研究。     

PAP-H2O2-HRP伏安酶联免疫分析新体系测定人血清总甲状腺素

目前临床检测中测定总甲状腺素(T₄)的常用方法有间接血凝试验、琼脂双扩散及ELISA等方法^[1].其中ELISA法是目前较为流行的检测方法,但灵敏度不高.伏安酶联免疫分析法具有广阔的应用前景^[2,3].     

基于金银纳米立方体构建高灵敏CA-153传感器的研究

癌抗原-153(CA-153)是乳腺癌最重要的特异性标志物。利用CA-153与其抗体之间的特异性识别性构建"三明治"夹心结构的免疫传感器,在玻碳电极上修饰金纳米/氧化石墨烯复合材料,通过纳米金和CA-153抗体之间的吸附作用,将抗体固定于电极表面,以牛血清白蛋白封闭非特异性吸附位点。金银(AuAg)纳米立方体标记CA-153二抗,标记的AuAg纳米立方体催化过氧化氢氧化电子媒介体硫堇,采用差分脉冲伏安法检测CA-153的电化学信号。在最优条件下,此传感器的响应电流与CA-153浓度的对数在2.0×10~(-5)~100 U/mL范围内呈良好的线性关系,检出限(S/N=3)为7.0×10~(-6)U/mL。对实际血清样品进行加标回收实验,回收率为92.2%~110.2%,相对标准偏差不大于8.7%。     

Differential pulse voltammetric enzyme-linked immunoassay for the determination of Helicobacter pylori specific immunoglobulin G (IgG) antibody

A differential pulse voltammetric enzyme-linked immunoassay for the determination of helicobacter pylori (H. pylori) specific IgG antibody in human serum has been developed. The method is based on coupling the oxidation reaction of 3,3',5,5'-tetramethylbenzidine (TMB)-H(2)O(2) that is catalysed by horseradish peroxidase-IgG(HRP-IgG) conjugate with the electro-reduction of the enzymatic product to measure the activity of HRP-IgG. The latter reaction exhibits a sensitive differential pulse voltammetric response at 0.1 V (versus Ag AgCl ) in pH 4.0 acetate buffer solution. So, the H. pylori specific IgG antibody could be detected. The detection limit of present method for H. pylori specific IgG antibody was 1.0 units ml(-1), which was about seven times lower than that obtained by traditional spectrophotometric ELISA procedure.     

Electrochemical studies of 1,5-dihydroxynaphthalene as substrate for voltammetric enzyme immunoassay

Immunoassay is one of the biochemical analytical techniques using the specific antigen antibody com-plexation for analytical purposes. It has extensive ap-plication in clinical diagnostics, prevention and cure of diseases, and virus diagnostics. The presentation and progress of immunoassay methodology are one of the greatest achievements of bioanalytical chemistry. It is estimated that several-hundred millions of immuno-analytical determinations are carried out every year all over the world. E…     

OAP-H~2O~2-HRP伏安酶联免疫分析新体系测定人血清铁蛋白

首次提出邻氨基酚(OAP)-H~2O~2-辣根过氧化物酶(HRP)伏安酶联免疫分析新体系并用于人血清中铁蛋白的测定.本方法以线性扫描二阶导数伏安法栓测HRP催化H~2O~2氧化OAP的产物,用于游离HRP和各种HRP标记物的测定,灵敏度均高于经典的ELISA显色光度法.测定游HPR的线性范围为1.0x10^-^1^2-4.0x10^-^9g/mL,检测限达6.0x10^-^1^3g/mL.本法对铁蛋白测定的线性范围为0.2-320ng/mL,用所建立的方法对人血清样品进行了测定,并与现行的ELISA显色光度法进行对照,二者相关性很好.对此伏安酶联免疫分析新体系的电极还原过程也进行了详细的研究.     

Fluorescence immunoassay of alpha-fetoprotein with iron(III) tetrasulfonatophthalocyanine as a mimetic enzyme labeling reagent

A new fluorimetric immunoassay for alpha-fetoprotein (AFP) has been developed using a novel promising mimetic peroxidase, iron(III) tetrasulfonatophthalocyanine (FeTSPc), as a labeling reagent to catalyze the fluorescence reaction of P- hydroxyphenylacetic acid (P-HPA) and hydrogen peroxide (H2O2). In the competitive immunoassay, anti-AFP antibody was coated on a 96-well plate (polystyrene) and a constant amount of FeTSPc-labeled AFP and a known amount of test solution were added. Non-labeled and FeTSPc-labeled AFP compete for binding to the plate-bound antibody. After the immunoreaction, the immunochemically adsorbed FeTSPc-AFP conjugate moiety was determined by measuring the fluorescence produced in a solution containing P-HPA and H2O2. AFP can be determined in the concentration range of 1-300 ng mL(-1) with a detection limit of 0.5 ng mL(-1).