Basic Research Laboratory, Department of Food Science and Technology and GreenTech-based Food Safety Research Group, BK21 Four, Chung-Ang University, Anseong, 17546, Republic of Korea.
School of Chemical Engineering, Yeungnam University, 280 Daehak-ro, Gyeongsan, Gyeongbuk, 38541, Republic of Korea.
Anal Chim Acta. 2024 Mar 22;1295:342287. doi: 10.1016/j.aca.2024.342287. Epub 2024 Jan 30.
This paper reports the development of a highly sensitive and selective electrochemical peptide-based biosensor for the detection of the inflammatory disease biomarker, interleukin-1beta (IL-1β). To this end, flower-like Au-Ag@MoS-rGO nanocomposites were used as the signal amplification platform to achieve a label-free biosensor with a high sensitivity and selectivity. First, a high-affinity peptide for IL-1β was identified through biopanning with M13 random peptide libraries, and was newly designed by incorporating cysteine at the C-terminus. An IL-1β specific binding peptide was used as the bio-receptor, and the interaction between the IL-1β binding peptide and IL-1β was confirmed via enzyme-linked immunosorbent assay and various physicochemical and electrochemical analyses. Under optimal conditions, the biosensor achieved an ultrasensitive and specific IL-1β detection in a wide linear concentration range of 0-250 ng/mL with a picomolar-level detection limit (∼2.4 pM), low binding constant (∼0.62 pM), and a low coefficient of variation (<1.65 %). The biosensor was successfully utilized for IL-1β determination in the serum of Crohn's disease patients with a good correlation coefficient. In addition, the detection performance was comparable to that of commercially available IL-1β ELISA kit. This indicates that the electrochemical peptide-based biosensor may offer a potentially valuable platform for the clinical diagnosis of various inflammatory disease biomarkers.
本文报道了一种基于肽的高灵敏度和选择性电化学生物传感器的开发,用于检测炎症性疾病生物标志物白细胞介素-1β(IL-1β)。为此,采用花状 Au-Ag@MoS-rGO 纳米复合材料作为信号放大平台,实现了具有高灵敏度和选择性的无标记生物传感器。首先,通过 M13 随机肽文库的生物淘选,鉴定出对 IL-1β具有高亲和力的肽,并通过在 C 末端掺入半胱氨酸对其进行新设计。IL-1β 特异性结合肽被用作生物受体,通过酶联免疫吸附测定和各种物理化学和电化学分析证实了 IL-1β 结合肽与 IL-1β 之间的相互作用。在最佳条件下,该生物传感器在 0-250ng/mL 的宽线性浓度范围内实现了超灵敏和特异性的 IL-1β 检测,检测限低至皮摩尔级(约 2.4pM),结合常数低(约 0.62pM),变异系数低(<1.65%)。该生物传感器成功地用于克罗恩病患者血清中 IL-1β 的测定,与商业可用的 IL-1β ELISA 试剂盒具有良好的相关性。这表明基于电化学肽的生物传感器可能为各种炎症性疾病生物标志物的临床诊断提供一个有价值的平台。