AbstractThe authors describe the fabrication of an electrochemical immunosensor for the determination of the activity of protein kinase A (PKA). The method involves (a) electrochemical deposition of gold nanoparticles (AuNPs) on a glassy carbon electrode, (b) PKA-induced catalytic phosphorylation of serine, and (c) the use of phosphoserine antibody and horseradish peroxidase conjugated to IgG on gold nanoparticles (HRP-IgG-AuNPs). The use of AuNPs and HRP-IgG-AuNPs results in large amplification so that the method, at a typical working potential as low as 0.08 V (vs. SCE), has a linear range that extends from 0.1 to 50 activity units per mL, and the detection limit is 0.026 units per mL (at an S/N ratio of 3). The assay is selective (not the least due to a rather low working potential) and well reproducible. It may also be applied to screening for PKA inhibitors and to quantify the PKA activity in human cell lysates. ᅟ

中文翻译：

---

使用与辣根过氧化物酶偶联的 IgG 功能化金纳米粒子修饰的玻璃碳电极对蛋白激酶 a 的活性进行电流法测定

摘要作者描述了用于测定蛋白激酶 A (PKA) 活性的电化学免疫传感器的制造。该方法包括（a）金纳米颗粒（AuNPs）在玻碳电极上的电化学沉积，（b）PKA诱导的丝氨酸催化磷酸化，以及（c）在金纳米颗粒上使用与IgG偶联的磷酸丝氨酸抗体和辣根过氧化物酶（ HRP-IgG-AuNPs）。AuNPs 和 HRP-IgG-AuNPs 的使用导致大的放大，因此该方法在低至 0.08 V（相对于 SCE）的典型工作电位下具有从 0.1 到 50 活性单位/mL 的线性范围，检测限为每毫升 0.026 个单位（信噪比为 3）。该测定具有选择性（尤其是由于工作潜力相当低）且重现性良好。它还可以用于筛选 PKA 抑制剂和量化人类细胞裂解物中的 PKA 活性。ㅟ