Porous reduced graphene oxide modified electrodes for the analysis of protein aggregation. Part 2: Application to the analysis of calcitonin containing pharmaceutical formulation
Porous reduced graphene oxide modified electrodes for the analysis of protein aggregation. Part 2: Application to the analysis of calcitonin containing pharmaceutical formulation
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DOI：
10.1016/j.electacta.2018.02.038
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发表时间：
2018-03-10
影响因子：
6.6
通讯作者：
Szunerits，Sabine
中科院分区：
文献类型：
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作者：
Vasilescu，Alina;Ye，Ran;Szunerits，Sabine
在第1部分（A. Vasilescu等人，多孔还原氧化石墨烯修饰电极用于蛋白质聚集分析。第1部分：pH 2和7.4电化学条件下的溶菌酶聚集。Acta，254（2017）375-383），我们提出了多孔还原氧化石墨烯涂覆的玻璃碳电极（GC/prGO）与差分脉冲伏安法的组合作为用于蛋白质聚集研究的新分析工具。溶菌酶被用作一个模型，通过电化学手段在pH 2和pH 7.4，导致淀粉样蛋白和无定形聚集体的形成，分别遵循其聚集。这项工作的第2部分通过研究生物制药产品扩大了这种方法的范围，因为基于肽的药物的聚集会影响其治疗活性，并可能诱导患者的过敏反应。鲑鱼多肽降钙素（sCT）被选择作为具有有限药物潜力的生物活性肽的实例，这是由于其倾向于形成细胞毒性聚集体和淀粉样蛋白原纤维。在实际应用中，采用逐层沉积技术制备了聚二烯丙基二甲基铵（PDDA）和prGO修饰的丝网印刷电极（SPE）和柔性电极（FE），用于sCT的检测。结果表明，这些电极可以区分在柠檬酸盐缓冲液中降钙素（2 mg mL-1）的淀粉样蛋白聚集体的形成与在乙酸盐缓冲液中无聚集。进一步证明，这些电极能够分析低效力的药物制剂Miacalcic（8.3 μg mL-1），其中未观察到聚集。
In part 1 (A. Vasilescu et al., Porous reduced graphene oxide modified electrodes for the analysis of protein aggregation. Part 1: Lysozyme aggregation at pH 2 and 7.4 Electrochem. Acta, 254 (2017) 375–383) we proposed porous reduced graphene oxide coated glassy carbon electrode (GC/prGO) in combination with differential pulse voltammetry as a new analytical tool for aggregation studies of proteins. Lysozyme was used as a model to follow its aggregation by electrochemical means at pH 2 and pH 7.4, leading to the formation of amyloid and amorphous aggregates, respectively. Part 2 of this work widens the scope of this approach by investigating a biopharmaceutical product, as the aggregation of peptide based drugs affects their therapeutic activity and can induce allergic reactions in patients. The salmon polypeptide calcitonin (sCT) was chosen as an example of a bioactive peptide with limited pharmaceutical potential due to a tendency to form cytotoxic aggregates and amyloid fibrils. For practical applications, screen printed electrodes (SPE) and flexible electrodes (FE) modified with polydiallyldimethylammonium (PDDA) and prGO by using the layer-by-layer deposition technique have been developed for the detection of sCT. The results indicate that these electrodes can differentiate between formation of amyloid aggregates of calcitonin (2 mg mL−1) in citrate buffer to no aggregation in acetate buffer. It was further demonstrated that these electrodes are able to analyze a pharmaceutical drug product of low potency, Miacalcic (8.3 μg mL−1), where no aggregation was observed.