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A Fiber Optic Interface Coupled to Nanosensors: Applications to Protein Aggregation and Organic Molecule Quantification.
ACS Nano ( IF 15.8 ) Pub Date : 2020-07-15 , DOI: 10.1021/acsnano.0c03417 Daichi Kozawa 1 , Soo-Yeon Cho 1 , Xun Gong 1 , Freddy T Nguyen 1 , Xiaojia Jin 1 , Michael A Lee 1 , Heejin Lee 2 , Alicia Zeng 2 , Gang Xue 2 , Jeff Schacherl 2 , Scott Gibson 2 , Leonela Vega 2 , Michael S Strano 1
ACS Nano ( IF 15.8 ) Pub Date : 2020-07-15 , DOI: 10.1021/acsnano.0c03417 Daichi Kozawa 1 , Soo-Yeon Cho 1 , Xun Gong 1 , Freddy T Nguyen 1 , Xiaojia Jin 1 , Michael A Lee 1 , Heejin Lee 2 , Alicia Zeng 2 , Gang Xue 2 , Jeff Schacherl 2 , Scott Gibson 2 , Leonela Vega 2 , Michael S Strano 1
Affiliation
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Fluorescent nanosensors hold promise to address analytical challenges in the biopharmaceutical industry. The monitoring of therapeutic protein critical quality attributes such as aggregation is a long-standing challenge requiring low detection limits and multiplexing of different product parameters. However, general approaches for interfacing nanosensors to the biopharmaceutical process remain minimally explored to date. Herein, we design and fabricate a integrated fiber optic nanosensor element, measuring sensitivity, response time, and stability for applications to the rapid process monitoring. The fiber optic–nanosensor interface, or optode, consists of label-free nIR fluorescent single-walled carbon nanotube transducers embedded within a protective yet porous hydrogel attached to the end of the fiber waveguide. The optode platform is shown to be capable of differentiating the aggregation status of human immunoglobulin G, reporting the relative fraction of monomers and dimer aggregates with sizes 5.6 and 9.6 nm, respectively, in under 5 min of analysis time. We introduce a lab-on-fiber design with potential for at-line monitoring with integration of 3D-printed miniaturized sensor tips having high mechanical flexibility. A parallel measurement of fluctuations in laser excitation allows for intensity normalization and significantly lower noise level (3.7 times improved) when using lower quality lasers, improving the cost effectiveness of the platform. As an application, we demonstrate the capability of the fully integrated lab-on-fiber system to rapidly monitor various bioanalytes including serotonin, norepinephrine, adrenaline, and hydrogen peroxide, in addition to proteins and their aggregation states. These results in total constitute an effective form factor for nanosensor-based transducers for applications in industrial process monitoring.
中文翻译：
与纳米传感器耦合的光纤接口：在蛋白质聚集和有机分子定量中的应用。
荧光纳米传感器有望解决生物制药行业中的分析难题。监视治疗性蛋白质的关键质量属性（例如聚集）是一项长期的挑战，需要低检测限和不同产品参数的复用。然而，迄今为止仍很少探索将纳米传感器与生物制药过程连接的一般方法。本文中，我们设计并制造了集成的光纤纳米传感器元件，可测量灵敏度，响应时间和稳定性，以用于快速过程监控。光纤-纳米传感器接口或光电二极管由无标签的nIR荧光单壁碳纳米管换能器组成，该换能器嵌入与光纤波导末端相连的保护性多孔水凝胶中。该光电二极管平台显示出能够区分人免疫球蛋白G的聚集状态，报告了在5分钟的分析时间内，分别具有5.6和9.6 nm大小的单体和二聚体聚集体的相对分数。我们引入了具有在线监测潜力的光纤实验室设计，并集成了具有高机械灵活性的3D打印的微型传感器头。当使用质量较低的激光器时，可以并行测量激光器激发的波动，从而实现强度归一化并显着降低噪声水平（提高了3.7倍），从而提高了平台的成本效益。作为一项应用，我们展示了完全集成的光纤实验室系统能够快速监测各种生物分析物的能力，包括5-羟色胺，去甲肾上腺素，肾上腺素和过氧化氢，除了蛋白质及其聚集状态。这些结果总体上构成了基于纳米传感器的换能器在工业过程监控中的有效形式。
更新日期：2020-08-25
中文翻译：
与纳米传感器耦合的光纤接口：在蛋白质聚集和有机分子定量中的应用。
荧光纳米传感器有望解决生物制药行业中的分析难题。监视治疗性蛋白质的关键质量属性（例如聚集）是一项长期的挑战，需要低检测限和不同产品参数的复用。然而，迄今为止仍很少探索将纳米传感器与生物制药过程连接的一般方法。本文中，我们设计并制造了集成的光纤纳米传感器元件，可测量灵敏度，响应时间和稳定性，以用于快速过程监控。光纤-纳米传感器接口或光电二极管由无标签的nIR荧光单壁碳纳米管换能器组成，该换能器嵌入与光纤波导末端相连的保护性多孔水凝胶中。该光电二极管平台显示出能够区分人免疫球蛋白G的聚集状态，报告了在5分钟的分析时间内，分别具有5.6和9.6 nm大小的单体和二聚体聚集体的相对分数。我们引入了具有在线监测潜力的光纤实验室设计，并集成了具有高机械灵活性的3D打印的微型传感器头。当使用质量较低的激光器时，可以并行测量激光器激发的波动，从而实现强度归一化并显着降低噪声水平（提高了3.7倍），从而提高了平台的成本效益。作为一项应用，我们展示了完全集成的光纤实验室系统能够快速监测各种生物分析物的能力，包括5-羟色胺，去甲肾上腺素，肾上腺素和过氧化氢，除了蛋白质及其聚集状态。这些结果总体上构成了基于纳米传感器的换能器在工业过程监控中的有效形式。
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