OAP-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H2O 2 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-0.87 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αFP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。     

ODA-H_2O_2-HRP伏安酶联免疫分析新体系的研究

提出邻联茴香胺-H_2O_2-HRP伏安酶联免疫分析新体系,并用于测定HRP和HRP标记物.该方法是将HRP催化H_2O_2氧化邻联茴香胺的酶催化反应与邻联茴香胺的氧化产物的电极还原反应相偶合,在BR缓冲溶液中,在-0.56V(SCE)左右产生灵敏的极谱波.应用此极谱波测定HRP的检测限为3.7×10~(-12)g/mL,线性范围为1.O×1O~(-11)～2.0×10~(-9)g/mL.对邻联茴香胺-H_2O_2-HRP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进行了较详细的探讨.     

OPD-H2O2-HRP伏安酶联免疫分析体系酶催化反应的研究

应用电化学分析、高效液相色谱、紫外-可见光谱、红外光谱和核磁共振等技术对邻苯二胺(OPD)-H2O2-辣根过氧化物酶(HRP)伏安酶联免疫分析体系的酶催化反应进行了详细深入的研究.用化学方法制得了HRP酶催化H2O2氧化OPD的产物纯品.伏安法和高效液相色谱实验说明,在所选择的酶催化反应条件下,酶催化反应只生成一种产物;经紫外-可见光谱,红外光谱和13C核磁共振谱鉴定,产物为2,3-二氨基吩嗪.写出了酶催化反应过程,同时对酶催化反应产物的电极还原过程也进行了研究.     

MAP-H2O2-HRP伏安酶联免疫分析新体系的光谱及电化学研究

提出了间氨基酚(MAP)-H2O2-辣根过氧化物酶(HRP)伏安酶联免疫分析新体系.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1.0×10-8～1.0×10-6g/L,检测限达3.8×10-9g/L.制备出了HRP催化H2O2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,13C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-羟苯基)氨基]-2,5-环己二烯基-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.     

甲基红-H2O2-HRP伏安酶联免疫分析法测定HRP及其标记物

提出了一种新的辣根过氧化物酶的底物-甲基红,它本身具有电化学活性,能够在静汞电极上发生还原反应,产生灵敏的伏安电流信号.以H2O2为氧化剂,HRP能催化氧化还原反应的发生,使甲基红被氧化分解,其平衡浓度降低,对应的还原峰电流降低,峰电流的降低值与HRP的质量浓度在5.0×10-8～5.0×10-7g/mL之间呈线性关系,对2.0×10-7g/mL HRP进行11次测定的相对标准偏差为4.6%,方法的检出限为1.8×10-8g/mL.应用于IgG-HRP和Avidin-HRP的测定.     

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.     

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     

MAP-H2O2-HRP伏安酶联免疫分析测定南方菜豆花叶病毒

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,并用于南方菜豆花叶病毒(SBMV)的测定.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及SBMV的测定,灵敏度均高于经典的ELISA显色光度法.本法对HRP测定的线性范围为1.0×10^-8～1.0×10^-6g/L,检测限为3.8×10^-9g/L;对SBMV测定的线性范围为4.0～5000ng/mL,检测限为4.0ng/mL.用所建立的方法测定病毒感染病叶澄清液的最高稀释比为1∶1.5×10⁵,并与现行的ELISA显色光度法进行对照,二者相关性很好.     

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].     

酸性铬蓝K-H2O2-HRP伏安酶联免疫分析法测定HRP及其标记物

本文提出了一种新的辣根过氧化物酶(HRP)的底物--酸性铬蓝K(ACBK),它本身具有电化学活性,能够在汞电极上发生还原反应.以H2O2为氧化剂,HRP的加入能加快氧化反应的进行,使酸性铬蓝K被氧化分解,其平衡浓度降低,对应的还原峰电流降低,峰电流的降低值同HRP的加入量在8.0×10-8～1.0×10-6 g/mL之间呈线性关系.用于IgG-HRP的测定,最高稀释比为1∶5 000.     

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.     

Investigation of voltammetric enzyme-linked immunoassay based on new system of ODA-H_2O_2-HRP

A voltammetric enzyme-linked immunoassay based on a new system of ODA-H2O2-HRP has first been developed and used in the detection of HRP and labelled HRP. By this method, the enzyme-catalyzing reaction of H2O2 oxidizing odianisidine (ODA) couples the electrode-reduction reaction of the oxidizing product of odianisidine, which produces a sensitive polarographic wave at potential of -0.56V (SCE) in Britton-Robinson buffer solution. In using this polarographic wave, a detection limit to HRP is 3.7×10-12g/mL and a linear range 1.0×10-11-2.0×10-9g/mL. And the mechanisms of the coupling reaction and the process of electro-reduction in the ODA-H2O2-HRP voltammetric enzyme-linked immunoassay system have also been carefully studied.     

偶合反应化学发光酶免疫分析研究: I. HRP及其标记物的化学发 光测定

本文将辣根过氧化物酶(HRP)催化过氧化氢氧化KI生成I~2的反应与Luminol-I~2的化学发光反应相偶合,提出了测定HRP及其抗原、抗体标记物的高灵敏度的化学发光法。检测下限为7pg,线性范围为10-6000pg。通常使用的HRP直接催化Luminol-H~2O~2化学发光法灵敏度高10-100倍;克服了固相吸附法直接测定HRP标记物的缺陷,提高了测定的选择性。文中还对该偶合反应的机理及其动力学特性进行了研究。     

电沉积CuInSe2上H2O2阴极还原时的电化学振荡行为

用电沉积方法得到的CuInSe2薄膜在阴极还原H2O2时发现了周期性的电化学振荡现象, 并研究了极化电位对该振荡和行为的影响。循环伏安测试表明电流-电势曲线中存在"电流波", 电流突变区域具有负斜率性质。用交流阻抗法研究了振荡体系的阻抗变化, 发现在振荡电势区域体系存在负电阻和电感成分, 这反映出振荡机一是中可能存在吸附中间物和自催化反应。     

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…     

Tl^3^+─H2O2─Fe^2^+体系的反应动力学研究

本文用Tl(I)离子选择性电极测定了Tl^3^+-H2O2-Fe^2^+体系的反应参数, 据此计算了该体系的反应级数, 得到了反应速率方程式, 对该反应体系的反应机理进行了探讨, 并用稳态处理法对所提出机理进行验证, 其结论与实验所得速率方程完全符合。     

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     

Detection of TMV with ODA-H(2)O(2)-HRP voltammetric enzyme-linked immunoassay system

o-Dianisidine (ODA)-H₂O₂-horseradish peroxidase (HRP) voltammetric enzyme-linked immunoassay system has firstly been used for the detection of tobacco mosaic virus (TMV). HRP catalyzes strongly the oxidation reaction of ODA by H₂O₂, the product of which produces a sensitive second order derivative linear sweep voltammetric peak at potential of −0.56 V (versus SCE) in Britton–Robinson (BR) buffer. HRP activity has been measured with this voltammetric peak and TMV detected through immunoreaction. The detection limit for HRP is 9.25×10^-7 mU l⁻¹ and the linear range is 2.5×10⁻⁶–5.0×10⁻⁴ mU l⁻¹. The detection limit for the clarified TMV is 0.25 ng ml⁻¹ and the highest dilution ratio detected for the infected leaf sap is 1:8×10⁵. The sensitivity for TMV detection with this method is higher than that with the enzyme-linked immunosorbent spectrophotometric assay (ELISA) using ODA-H₂O₂-HRP system. The processes of the enzyme-catalyzed reaction and the electro-reduction of the product of the enzyme-catalyzed reaction have been described.     

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。