Biosensor National Special Laboratory, Key Laboratory for Biomedical Engineering of Education Ministry, Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, China.
Cancer Centre, Zhejiang University, Hangzhou 310058, China.
Biosensors (Basel). 2022 Aug 9;12(8):619. doi: 10.3390/bios12080619.
Conventional immunocolorimetric sensing of biomolecules continues to be challenged with low sensitivity although its wide application as a diagnostic tool in medicine and biotechnology. Herein, we present a multifunctional immunocolorimetric sensing system for sensitive and high-throughput detection of B-type natriuretic peptide (BNP) with carbon-gold nanocomposite (CGNs) amplification. Using a "green" strategy, monodisperse carbon nanospheres (CNs) were successfully synthesized by glucose carbonization. A simple and efficient hydrothermal method was developed to assemble abundant gold nanoparticles (AuNPs) onto the surfaces of CNs. The resulting CGNs were characterized and utilized for biomarker detection with superior properties of easy manufacturing, good biocompatibility, satisfactory chemical stability, and high loading capacity for biomolecules. As a proof of concept, the as-prepared CGNs were conjugated with horseradish peroxidase-labeled antibody against BNP (CGNs@AntiBNP-HRP) functioning as the carrier, signal amplifier, and detector for the sensitive detection of BNP. Under optimal conditions, the established CGNs@AntiBNP-HRP immunoprobe remarkably enhanced the detection performance of BNP, achieving signal amplification of more than 9 times compared to the conventional method. Based on our self-developed bionic electronic eye (e-Eye) and CGNs@AntiBNP-HRP immunoprobe, the multifunctional sensing system displayed a wide dynamic linear range of 3.9-500 ng/mL and a LOD of 0.640 ng/mL for BNP detection with high specificity, good accuracy and reproducibility. This portable sensing system with enhanced performance demonstrates great potential for BNP detection in point of care applications, and offers a universal and reliable platform for in vitro biomarker detection.
尽管传统的免疫比色传感在医学和生物技术中的诊断工具方面得到了广泛应用，但由于其灵敏度较低，仍然面临挑战。在此，我们提出了一种多功能免疫比色传感系统，用于灵敏和高通量检测 B 型利钠肽（BNP），该系统采用碳-金纳米复合材料（CGNs）进行放大。利用“绿色”策略，通过葡萄糖碳化成功合成了单分散的碳纳米球（CNs）。开发了一种简单高效的水热法，将丰富的金纳米颗粒（AuNPs）组装到 CNs 的表面上。所得 CGNs 进行了表征，并用于生物标志物检测，具有易于制造、良好的生物相容性、令人满意的化学稳定性和高生物分子负载能力等优异性能。作为概念验证，将制备的 CGNs 与辣根过氧化物酶标记的针对 BNP 的抗体（CGNs@AntiBNP-HRP）偶联，作为载体、信号放大器和检测器，用于 BNP 的灵敏检测。在最佳条件下，所建立的 CGNs@AntiBNP-HRP 免疫探针显著提高了 BNP 的检测性能，与传统方法相比，信号放大超过 9 倍。基于我们自主开发的仿生电子眼（e-Eye）和 CGNs@AntiBNP-HRP 免疫探针，多功能传感系统显示出宽的动态线性范围为 3.9-500ng/mL，LOD 为 0.640ng/mL，对 BNP 检测具有高特异性、良好的准确性和重现性。该具有增强性能的便携式传感系统在即时护理应用中具有很大的 BNP 检测潜力，并为体外生物标志物检测提供了通用可靠的平台。
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