da Silva Jordana S L, Oliveira Maria D L, de Melo Celso P, Andrade Cesar A S
Programa de Pós-Graduação em Saúde Humana e Meio Ambiente, UFPE, 55608-680 Vitória de Santo Antão, PE, Brazil.
Departamento de Bioquímica, UFPE, 50670-901 Recife, PE, Brazil.
Colloids Surf B Biointerfaces. 2014 May 1;117:549-54. doi: 10.1016/j.colsurfb.2013.12.057. Epub 2014 Jan 6.
In this paper, we report the use of Concanavalin A (ConA) and electrosynthesized polyaniline (PANI) thin films for the development of a new electrochemical sensor that allows the specific detection of two bacterial toxins: lipopolysaccharide (LPS) from Escherichia coli and lipoteichoic acid from Staphylococcus aureus. The impedimetric sensor is fabricated by using glutaraldehyde to self-assemble ConA lectin on PANI-modified steel electrodes through covalent binding. ConA acts as a recognition element for bacterial toxins. Electrical impedance spectroscopy (EIS) and scanning electron microscope (SEM) were applied to characterize the assembly process on the modified electrode. The EIS measurements revealed that the resistance charge transfer (RCT) of the electrode/electrolyte interface increases considerably after the ConA lectin interacts with specific carbohydrate moieties present in the molecule of the bacterial toxin. Our results showed that the ConA lectin retained its activity after immobilization on the PANI surface and also the existence of electrochemical impedance response of the bioelectrode which is linear to the extent of the lectin-toxin interaction, with maximum values of RCT for E. coli (14.40 kΩ), and S. aureus (17.80 kΩ). We have observed that electrosynthesized PANI is an excellent support layer for the covalent binding of lectins on the electrode surface. Thus, the recognition system provides an appropriate biomimetic interface for detection of specific constituents of gram-positive and gram-negative bacteria.
在本文中,我们报道了使用伴刀豆球蛋白A(ConA)和电合成聚苯胺(PANI)薄膜开发一种新型电化学传感器,该传感器能够特异性检测两种细菌毒素:大肠杆菌的脂多糖(LPS)和金黄色葡萄球菌的脂磷壁酸。通过使用戊二醛将ConA凝集素通过共价结合自组装在PANI修饰的钢电极上,制备了阻抗传感器。ConA作为细菌毒素的识别元件。应用电阻抗谱(EIS)和扫描电子显微镜(SEM)对修饰电极上的组装过程进行表征。EIS测量结果表明,在ConA凝集素与细菌毒素分子中存在的特定碳水化合物部分相互作用后,电极/电解质界面的电阻电荷转移(RCT)显著增加。我们的结果表明,ConA凝集素固定在PANI表面后仍保留其活性,并且生物电极存在电化学阻抗响应,该响应与凝集素 - 毒素相互作用的程度呈线性关系,大肠杆菌的RCT最大值为14.40 kΩ,金黄色葡萄球菌的RCT最大值为17.80 kΩ。我们观察到电合成的PANI是凝集素在电极表面共价结合的优良支撑层。因此,该识别系统为检测革兰氏阳性和革兰氏阴性细菌的特定成分提供了合适的仿生界面。