Enzyme sensor for the electrochemical detection of the marine toxin okadaic acid
Enzyme sensor for the electrochemical detection of the marine toxin okadaic acid
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DOI：
10.1016/j.aca.2007.10.036
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发表时间：
2007-12-12
影响因子：
6.2
通讯作者：
Marty, Jean-Louis
中科院分区：
文献类型：
--
作者：
Campas, Monica;Marty, Jean-Louis
研制了一种用于海洋毒素冈田酸（OA）电化学检测的酶传感器。该策略是基于这种毒素对固定化蛋白磷酸酶（PP2A）的抑制，以及通过使用适当的酶底物对酶活性的电化学测量，酶在去磷酸化后具有电化学活性。比色法抑制试验表明，来自人红细胞的PP2A比基因工程产生的PP2A更敏感，并提供更宽的线性范围。单磷酸儿茶酚（CMP）和对氨基苯基磷酸（PAPP）作为酶底物进行了测试，前者在方便的工作电位下（+450 mV us）提供较高的电化学电流。Ag / AgCl)。将固定化酶浓度分别为19.1和5.0 U的生物传感器应用于OA检测。与19.1-U生物传感器提供更高的电化学电流和更可靠的测定相比，5.0-U生物传感器具有更低的50%抑制系数（IC50）值（在154.84 mu g -1前为22.19）和更大的工作范围（在42.97-171.87 mu g -1前为2.69-171.87）。利用生物传感器对有毒的鞭毛藻提取物进行分析，并与比色法和液相色谱串联质谱法（LC-MS/MS）进行比较，证明了所开发的电化学装置作为评估样品毒性的筛选生物工具的适用性。(c) 2007 Elsevier B.V.版权所有
An enzyme sensor for the electrochemical detection of the marine toxin okadaic acid (OA) has been developed. The strategy was based on the inhibition of immobilised protein phosphatase (PP2A) by this toxin and the electrochemical measurement of the enzyme activity by the use of appropriate enzyme substrates, electrochemically active after dephosphorylation by the enzyme. Colorimetric inhibition assays have demonstrated the PP2A from human red blood cells to be more sensitive and to provide a wider linear range than the one produced by genetic engineering. Catechyl monophosphate (CMP) and p-aminophenyl phosphate (PAPP) have been tested as enzyme substrates, the former providing higher electrochemical currents at convenient working potentials (+450 mV us. Ag/AgCl). Biosensors with 19.1 and 5.0 U of immobilised enzyme have been applied to the OA detection. Whereas the 19.1-U biosensor has provided higher electrochemical currents and more reliable determinations, the 5.0-U one has attained a lower 50% inhibition coefficient (IC50) value (22.19 in front of 154.84 mu g L-1) and a larger working range (2.69-171.87 in front of 42.97-171.87 mu g L-1). The analysis of toxicogenic dinoflagellate extracts with both biosensors and the comparison with the colorimetric assay and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) have demonstrated the applicability of the developed electrochemical devices as screening biotools for the assessment of the toxicity of a sample. (c) 2007 Elsevier B.V. All rights reserved.