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     

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。     

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     

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。     

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）     

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     

联苯胺—H2O2—HRP偶合反应伏安酶联免疫分析新体系测定HRP的研究

提出了联苯胺－Ｈ２Ｏ３－ＨＲＰ偶合反应伏安酶联免疫分析新体系测定ＨＲＰ的方法。通过测定ＨＲＰ催化Ｈ２Ｏ２氧化联苯胺的产物间接测定ＨＲＰ的浓度。     

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类风湿因子抗体

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所形成酶催化产物在ｐＨ２．０磷酸盐－枸橼酸缓冲溶液中于－０．１８ Ｖ左右产生一灵敏示差脉冲伏安峰，ＲＦ浓度在１．２５～２０．０ Ｕ／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显色光度测定法的灵敏度增加５倍，且受干扰较少。     

用OPD-H 2 O 2-HRP伏安酶联免疫法检测植物病毒ArMV、C MV、SBMV和TuMV*

首次将邻苯二胺（ＯＰＤ）－Ｈ２Ｏ２－辣根过氧化物酶（ＨＲＰ）伏安酶联免疫分析体系,应用于植物病毒南芥菜花叶病毒（ＡｒＭＶ）,黄瓜花叶病毒（ＣＭＶ）,南方菜豆花叶病毒（ＳＢＭＶ）和芜箐花叶病毒的血清学检测。该法测定以上植物病毒感染病叶澄清液的最高稀释比（体积比）分别为１：５００００００,１：５０００００;１：２５０００００和１：２５００００,而酶联免疫吸附分析（ＥＬＩＳＡ）测定的最高稀释比分别为１：     

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     

MAP-H~2O~2-HPR伏安酶联免疫分析新体系和光谱及电化学研究

提出了间氨基酸(MAP)-H~2O~2-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H~2O~2氧化MAP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.测定游离HRP的线性范围为1.0x10^-^8-1.0x10-6/L,检测限达3.8x10^-^9g/L.制备出了HRP催化H~2O~2氧化MAP的产物纯品并应用电化学分析,高效液相色谱,元素分析,紫外-可见光谱,红外光谱,^1H核磁共振谱,^1^3C核磁共振谱及质谱等技术对体系酶促反应进行了深入的研究.在选择的酶促反应条件下,生成的产物为2-氨基-5-[(3-差苯基)]-2,5-环己烯基-1,4-二酮.提出了酶催化反应机理及其产物的电极还原过程。     

An improved ELISA for the determination of tobacco mosaic virus with linear sweep voltammetry detection based on a new system of PAP-H2O2-HRP

An improved ELISA for the determination of tobacco mosaic virus (TMV) with linear sweep voltammetry based on a new system of p-aminophenol (PAP)- H₂O₂- horseradish peroxidase (HRP) has been developed. The enzymatic product 3-[(4-hydroxyphenyl)amino]-4-(2-amino-5-hydroxyphenyl)-6-[(4-hydroxyphenyl)imino]-2,4-cyclohexadiene-1-one, produced from HRP catalyzing the oxidation of PAP with H₂O₂, yields a sensitive linear sweep voltammetric response at a potential of –0.45 V (vs. SCE) in Britton-Robinson (BR) buffer solution. By using this voltammetric peak, HRP can be measured with a detection limit of 0.4 mU/L and a linear range of 1.0 ～ 1.0 × 10² mU/ L. The detection limit for the clarified TMV is 4.0 ng/mL and the highest dilution ratio detected for the infected leaf sap is 1?:?3.9 × 10⁶. The processes of the enzyme-catalyzed reaction and the electro-reduction of the product of the enzyme-catalyzed reaction have been investigated.     

An improved ELISA for the determination of tobacco mosaic virus with linear sweep voltammetry detection based on a new system of PAP-H2O2-HRP

An improved ELISA for the determination of tobacco mosaic virus (TMV) with linear sweep voltammetry based on a new system of p-aminophenol (PAP)- H₂O₂- horseradish peroxidase (HRP) has been developed. The enzymatic product 3-[(4-hydroxyphenyl)amino]-4-(2-amino-5-hydroxyphenyl)-6-[(4-hydroxyphenyl)imino]-2,4-cyclohexadiene-1-one, produced from HRP catalyzing the oxidation of PAP with H₂O₂, yields a sensitive linear sweep voltammetric response at a potential of –0.45 V (vs. SCE) in Britton-Robinson (BR) buffer solution. By using this voltammetric peak, HRP can be measured with a detection limit of 0.4 mU/L and a linear range of 1.0 ∼ 1.0 × 10² mU/ L. The detection limit for the clarified TMV is 4.0 ng/mL and the highest dilution ratio detected for the infected leaf sap is 1 : 3.9 × 10⁶. The processes of the enzyme-catalyzed reaction and the electro-reduction of the product of the enzyme-catalyzed reaction have been investigated. Received: 17 November 1998 / Revised: 17 March 1999 / Accepted: 20 March 1999     

Enzyme-catalyzed reaction of OPD-H_2O_2-HRP voltammetric enzyme-linked immunoassay system

Ｈｏｒｓｅｒａｄｉｓｈｐｅｒｏｘｉｄａｓｅ(ＨＲＰ)ｃａｎｓｔｒｏｎｇｌｙｃａｔａｌｙｚｅｔｈｅｒｅａｃｔｉｏｎｏｆＨ2Ｏ2ｏｘｉｄｉｚｉｎｇｏｐｈｅｎｙｌｅｎｅｄｉａｍｉｎｅ(ＯＰＤ),ｔｈｅｃｈｒｏｍａｔｉｃｐｒｏｄｕｃｔｏｆｗｈｉｃｈｃａｎｂｅｄｅｔｅｃｔｅｄｉｎｅｎｚｙｍｅｌｉｎｋｅｄｉｍｍｕｎｏｓｏｒｂｅｎｔａｓｓａｙ(ＥＬＩＳＡ)ｗｉｔｈｓｐｅｃｔｒｏｐｈｏｔｏｍｅｔｒｙ[1].Ｉｎｖｏｌｔａｍｍｅｔｒｉｃｅｎｚｙｍｅｌｉｎｋｅｄｉｍｍｕｎｏａｓｓａｙ[2—4],ＯＰＤＨ2Ｏ2ＨＲＰｓｙｓｔｅｍｈａｓｆ…     

Enzyme-catalyzed reaction of voltammetric enzyme-linked immunoassay system based on OAP as substrate

The o-aminophenol (OAP)-H_2O_2-horseradish peroxidase (HRP) voltammetric enzyme-linked immunoassay new system has extremely high sensitivity. HRP can be measured with a detection limit of 6.0×10~-(10) g/L and a linear range of 1.0×10~(-9)—4.0×10~(-6) g/L. The pure product of H_2O_2 oxidizing OAP catalyzed by HRP was prepared with chemical method. The enzyme-catalyzed reaction has been investigated with electroanalytical chemistry, UV/Vis spectrum, IR spectrum, ~(13)C NMR, ~1H NMR, mass spectrum, elemental analysis, etc. Under the selected enzyme-catalyzed reaction conditions, the oxidation product of OAP with H_2_O2 catalyzed by HRP is 2-aminophe-noxazine-3-one. The processes of the enzyme-catalyzed reaction and the electroreduction of the product of the enzymecatalyzed reaction have been described.     

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].     

Enzyme-catalyzed reaction of OPD-H202-HRP voltammetric enzyme-linked immunoassay system

Enzyme-catalyzed reaction of o-phenylenediamine (OPD)-H_z0₂-horseradish peroxidase (HRP) voltammetric enzyme-linked immunoassay system has been studied in detail with electrochemical analysis, high performance liquid chromatography (HPLC), ultraviolet/visible (UV/Vis) spectroscopy, infrared (IR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. The pure product of H₂0₂ oxidizing OPD catalyzed by HRP was prepared with chemical method. The experimental results of voltammetry and HPLC indicate that only one product of enzyme-catalyzed reaction has been obtained under the selected enzyme-catalyzed reaction conditions. Identifications by UV/ Vis spectrum, IR spectrum and¹³C NMR spectrum show that the product is 2,3-diaminophenazine. The processes of the enzyme-catalyzed reaction and the electroreduction of the product of the enzyme-catalyzed reaction are described. Project supported by the National Natural Science Foundation of China     

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.     

Detection of ferritin in human serum with a MAP-H(2)O(2)-HRP voltammetric enzyme-linked immunoassay system

A voltammetric enzyme-linked immunoassay based on new system of m-aminophenol (MAP)-H(2)O(2)-horseradish peroxidase (HRP) has firstly been developed and used for the detection of HRP, labelled HRP and ferritin in human serum. HRP or labelled HRP catalyzes the oxidation reaction of MAP with H(2)O(2), the product of which produces a sensitive voltammetric peak at potential of -0.46 V (vs. SCE) in Britton-Robinson (BR) buffer solution. By using this voltammetric peak, HRP can be measured with a detection limit of 9.5x10(-1) mU/l and a linear range of 2.5-2.5x10(2) mU/l. The detection limit to ferritin is 0.25 ng/ml and the linear range 0.25-320 ng/ml. The processes of the electro-reduction of the product of the enzyme-catalyzed reaction have been investigated in detail.     

新型碳苷糖类维生素D_2衍生物的合成

以D-葡萄糖为原料,经碳苷化反应,酰化反应和脯氨酸-DIPEA催化的aldol反应制得2个碳苷糖[1-(4'-羟基苯基)-4-C-β-四乙酰基葡萄糖基-3-烯-2-酮(5a)和1-(3-羟基苯基)-4-C-β-四乙酰基葡萄糖基-3-烯-2-酮(5b)];5与琥珀酸维生素D2经Steglich酯化反应合成了2个新型碳苷糖类维生素D2衍生物,其结构经1H NMR,13C NMR和HR-ESI-MS表征。