Luna Débora M N, Oliveira Maria D L, Nogueira Maurício L, Andrade Cesar A S
Programa de Pós-Graduação em Inovação Terapêutica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
Departamento de Bioquímica, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.
Chem Phys Lipids. 2014 May;180:7-14. doi: 10.1016/j.chemphyslip.2014.02.008. Epub 2014 Mar 11.
In this work, we developed a biosystem based on Concanavalin A (ConA) and lipid membranes to recognize glycoproteins from the serum of patients contaminated with dengue serotypes 1, 2 and 3 (DENV1, DENV2 and DENV3). The modified gold electrode was characterized using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and atomic force microscopy. Morphological analyses of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), DPPC-ConA, DPPC-ConA-DENV1, DPPC-ConA-DENV2 and DPPC-ConA-DENV3 revealed the existence of a non-uniform covering and large globules. EIS and CV measurements have shown that redox probe reactions on the modified gold electrodes were partially blocked due to the adsorption of lipid-ConA system and reveal the interaction response of the immobilized ConA to the presence of glycoproteins of dengue serum. The biosystem exhibited a wide linear response to different concentrations of sera of dengue serotypes 1, 2 and 3. A higher impedimetric response to glycoproteins present in dengue serotype 3 was observed. Our results demonstrate the applicability of lectin and lipid membranes to the development of biosensors for dengue infections.
在这项工作中,我们开发了一种基于伴刀豆球蛋白A(ConA)和脂质膜的生物系统,用于识别感染登革热血清型1、2和3(DENV1、DENV2和DENV3)患者血清中的糖蛋白。使用循环伏安法(CV)、电化学阻抗谱(EIS)和原子力显微镜对修饰的金电极进行了表征。对1,2-二棕榈酰-sn-甘油-3-磷酸胆碱(DPPC)、DPPC-ConA、DPPC-ConA-DENV1、DPPC-ConA-DENV2和DPPC-ConA-DENV3的形态分析揭示了存在不均匀覆盖和大颗粒。EIS和CV测量表明,由于脂质-ConA系统的吸附,修饰金电极上的氧化还原探针反应被部分阻断,并揭示了固定化ConA对登革热血清糖蛋白存在的相互作用响应。该生物系统对不同浓度的登革热血清型1、2和3血清表现出广泛的线性响应。观察到对登革热血清型3中存在的糖蛋白有更高的阻抗响应。我们的结果证明了凝集素和脂质膜在开发用于登革热感染的生物传感器方面的适用性。