High sensitivity lab-on-fiber biosensing platform assisted by oriented antibody immobilization strategy

S. Ucci, S. Spaziani, G. Quero, P. Vaiano, M. Principe, A. Micco, A. Sandomenico, M. Ruvo, M. Consales, A. Cusano
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Abstract

The demand for highly sensitive, fast and low-cost biosensors for reliable quantification of small biomolecules or cancer biomarkers is leading to the development of a new class of devices able to change the techniques currently used for diagnosis in oncology. Lab‐on‐fiber (LoF) optrodes offer several advantages over conventional techniques for point‐of‐care platforms aimed at real‐time and label‐free detection of clinically relevant biomarkers. Moreover, the easy integration of LoF platforms in medical needles, catheters and nano-endoscopes offers unique potentials for in vivo biopsies and tumor microenvironment assessment. Here, we demonstrate the capability to improve the immobilization strategies through the use of hinge carbohydrates by involving homemade antibodies that demonstrated a significantly improved recognition of the antigen with ultra‐low detection limits. In order to create an effective pipeline for the improvement of biofunctionalization protocols to be used in connection with the LoF platform, here we first, optimized the protocol using a microfluidic Surface Plasmon Resonance device. Then we transferred the optimized strategy on LoF platform, based on Optical Fiber Meta‐tip (OFMT), for the final validation. As a clinically relevant scenario, we focused on a serological biomarker, Cripto‐1, for its ability to promote tumorigenesis in breast and liver cancer. Reported results demonstrate that the proposed approach based on oriented antibody immobilization is able to significantly improve Cripto‐1 detection with a ten‐fold enhancement versus the random approach. Therefore, our work opens new avenues in the development of high‐sensitivity LoF biosensors for the detection of clinically relevant biomarkers in the sub‐ng/mL range.
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定向抗体固定化策略辅助的高灵敏度光纤实验室生物传感平台
对高灵敏度,快速和低成本的生物传感器的需求,以可靠地定量小生物分子或癌症生物标志物,导致了一类新型设备的开发,能够改变目前用于肿瘤诊断的技术。光纤上实验室(LoF)光极在实时和无标签检测临床相关生物标志物的护理点平台上提供了许多优于传统技术的优势。此外,将LoF平台轻松集成到医用针头、导管和纳米内窥镜中,为活体活检和肿瘤微环境评估提供了独特的潜力。在这里,我们展示了通过使用铰链碳水化合物来改善固定策略的能力,通过使用自制抗体,可以显著提高对抗原的识别,并且具有超低的检测限。为了建立一个有效的管道来改进与LoF平台相关的生物功能化方案,我们首先使用微流体表面等离子体共振装置优化了方案。然后,我们将优化策略转移到基于光纤元端(OFMT)的LoF平台上进行最终验证。作为临床相关的场景,我们关注了一种血清学生物标志物,Cripto‐1,因为它能够促进乳腺癌和肝癌的肿瘤发生。报告的结果表明,基于定向抗体固定的方法能够显著提高Cripto‐1的检测,比随机方法提高10倍。因此,我们的工作为开发用于检测亚微克/毫升范围内临床相关生物标志物的高灵敏度LoF生物传感器开辟了新的途径。
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