Nanoporous gold as a solid support for protein immobilization and development of an electrochemical immunoassay for prostate specific antigen and carcinoembryonic antigen.
Nanoporous gold as a solid support for protein immobilization and development of an electrochemical immunoassay for prostate specific antigen and carcinoembryonic antigen.
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DOI：
10.1007/s00604-012-0870-x
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发表时间：
2012-10-01
影响因子：
5.7
通讯作者：
Stine, Keith J.
中科院分区：
文献类型：
--
作者：
Pandey, Binod;Demchenko, Alexei V.;Stine, Keith J.
关键词：
利用纳米孔金（NPG）作为载体，固定化碱性磷酸酶（ALP）与针对前列腺特异性抗原（PSA）或癌胚抗原（CEA）的单克隆抗体结合。抗体- alp偶联物偶联到硫辛酸自组装单层上，并用于直接动力学分析。以对氨基苯基磷酸为底物，用方波伏安法在0.1 V （vs. Ag|AgCl）附近氧化检测产物对氨基酚。与生物标志物孵育前后对氨基酚氧化产生的峰值电流差异随着生物标志物浓度的增加而增加。这两种生物标志物对CEA的反应为10 ng mL-1，对PSA的反应为30 ng mL-1。以牛血清白蛋白（BSA）为模型白蛋白，研究了干扰对PSA测定的影响。干扰的影响，从血清基质检查为PSA测定使用新生牛犊血清。在较低的抗原浓度下，将抗原固定在NPG表面的竞争性免疫测定方法具有很高的敏感性。给出了抗体- alp偶联物在NPG表面覆盖的估计。
Nanoporous gold (NPG) was utilized as a support for immobilizing alkaline phosphatase (ALP) conjugated to monoclonal antibodies against either prostate specific antigen (PSA) or carcinoembryonic antigen (CEA). The antibody-ALP conjugates were coupled to self-assembled monolayers of lipoic acid and used in direct kinetic assays. Using the enzyme substrate p-aminophenylphosphate, the product p-aminophenol was detected by its oxidation near 0.1 V (vs. Ag|AgCl) using square wave voltammetry. The difference in peak current arising from oxidation of p-aminophenol before and after incubation with biomarker increased with biomarker concentration. The response to these two biomarkers was linear up to 10 ng mL-1 for CEA and up to 30 ng mL-1 for PSA. The effect of interference on the PSA assay was studied using bovine serum albumin (BSA) as a model albumin protein. The effect of interference from a serum matrix was examined for the PSA assay using newborn calf serum. A competitive version of the immunoassay using antigen immobilized onto the NPG surface was highly sensitive at lower antigen concentration. Estimates of the surface coverage of the antibody-ALP conjugates on the NPG surface are presented.
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影响因子：
1
作者：
Pandey B;Tan YH;Fujikawa K;Demchenko AV;Stine KJ
通讯作者：
Stine KJ
影响因子：
17.1
作者：
Mani, Vigneshwaran;Chikkaveeraiah, Bhaskara V.;Patel, Vyomesh;Gutkind, J. Silvio;Rusling, James F.
通讯作者：
Rusling, James F.
影响因子：
--
作者：
Li, Yan;Gao, Fei;Zhou, Wei-Qing
通讯作者：
Zhou, Wei-Qing
影响因子：
6.6
作者：
Djavan, B;Zlotta, AR;Marberger, M
通讯作者：
Marberger, M
影响因子：
3.4
作者：
Seker E;Reed ML;Begley MR
通讯作者：
Begley MR